MXPA98000499A - Inhibition of the expression of ge - Google Patents
Inhibition of the expression of geInfo
- Publication number
- MXPA98000499A MXPA98000499A MXPA/A/1998/000499A MX9800499A MXPA98000499A MX PA98000499 A MXPA98000499 A MX PA98000499A MX 9800499 A MX9800499 A MX 9800499A MX PA98000499 A MXPA98000499 A MX PA98000499A
- Authority
- MX
- Mexico
- Prior art keywords
- sequence
- nucleotide sequence
- enzyme
- present
- intron
- Prior art date
Links
- 230000014509 gene expression Effects 0.000 title abstract description 27
- 230000005764 inhibitory process Effects 0.000 title description 2
- 239000002773 nucleotide Substances 0.000 claims abstract description 112
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 112
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 56
- 238000000034 method Methods 0.000 claims abstract description 50
- 230000002255 enzymatic effect Effects 0.000 claims abstract description 23
- 210000000056 organ Anatomy 0.000 claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 claims description 54
- 102000004190 Enzymes Human genes 0.000 claims description 53
- 229920002472 Starch Polymers 0.000 claims description 49
- 235000019698 starch Nutrition 0.000 claims description 49
- 239000008107 starch Substances 0.000 claims description 41
- 102000004169 proteins and genes Human genes 0.000 claims description 23
- 230000000694 effects Effects 0.000 claims description 21
- 230000009261 transgenic effect Effects 0.000 claims description 17
- 230000000692 anti-sense effect Effects 0.000 claims description 16
- 108700024394 Exon Proteins 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 12
- 238000010276 construction Methods 0.000 claims description 10
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 8
- 230000002829 reductive effect Effects 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 87
- 239000013598 vector Substances 0.000 description 32
- 244000061456 Solanum tuberosum Species 0.000 description 26
- 239000013612 plasmid Substances 0.000 description 25
- 235000002595 Solanum tuberosum Nutrition 0.000 description 24
- 235000018102 proteins Nutrition 0.000 description 14
- 239000012634 fragment Substances 0.000 description 13
- 239000000758 substrate Substances 0.000 description 13
- 230000009466 transformation Effects 0.000 description 11
- 230000036961 partial effect Effects 0.000 description 9
- 230000002068 genetic effect Effects 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 102000003925 1,4-alpha-Glucan Branching Enzyme Human genes 0.000 description 5
- 108090000344 1,4-alpha-Glucan Branching Enzyme Proteins 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000007246 mechanism Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 4
- 108091092195 Intron Proteins 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 241000589158 Agrobacterium Species 0.000 description 3
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 3
- 241000723289 Dianella <angiosperm> Species 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 3
- 229920000881 Modified starch Polymers 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 238000012136 culture method Methods 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 235000019426 modified starch Nutrition 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 235000012015 potatoes Nutrition 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 240000004760 Pimpinella anisum Species 0.000 description 2
- 108010064851 Plant Proteins Proteins 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 235000021118 plant-derived protein Nutrition 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 108010013043 Acetylesterase Proteins 0.000 description 1
- 241000589156 Agrobacterium rhizogenes Species 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 108020000992 Ancient DNA Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 101100219344 Arabidopsis thaliana CAT7 gene Proteins 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241001432959 Chernes Species 0.000 description 1
- 241000207199 Citrus Species 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241001331845 Equus asinus x caballus Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 239000005980 Gibberellic acid Substances 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 108010060309 Glucuronidase Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 108010028688 Isoamylase Proteins 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000218922 Magnoliophyta Species 0.000 description 1
- 102100036617 Monoacylglycerol lipase ABHD2 Human genes 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102220513475 Rab-interacting lysosomal protein_E35S_mutation Human genes 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 102100032741 SET-binding protein Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 101100352756 Schizosaccharomyces pombe (strain 972 / ATCC 24843) pnu1 gene Proteins 0.000 description 1
- 101100528946 Schizosaccharomyces pombe (strain 972 / ATCC 24843) rpa1 gene Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 108700026226 TATA Box Proteins 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 1
- 229960003669 carbenicillin Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000020971 citrus fruits Nutrition 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000001952 enzyme assay Methods 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 102000034356 gene-regulatory proteins Human genes 0.000 description 1
- 108091006104 gene-regulatory proteins Proteins 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 150000004688 heptahydrates Chemical class 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 108091070501 miRNA Proteins 0.000 description 1
- 239000002679 microRNA Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- DHCDFWKWKRSZHF-UHFFFAOYSA-N sulfurothioic S-acid Chemical compound OS(O)(=O)=S DHCDFWKWKRSZHF-UHFFFAOYSA-N 0.000 description 1
- 229910021653 sulphate ion Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 1
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Abstract
A method for inhibiting the expression of genes is described, the method, which affects the enzymatic activity in a plant, comprises expressing in a plant (a cell, a tissue or an organ thereof) a sequence of nucleotides, wherein the sequence of nucleotides encode, partially or completely, for an intron in a sense orientation, and wherein the nucleotide sequence does not contain a sequence that is sense to an exon sequence normally associated with the intr
Description
INHIBITION OF LES EXPRESSION OF GENES
II ESCRIPTIVE ENVIRONMENT
The present invention relates to a method for inhibiting the expression of genes, particularly to inhibit the expression of genes in a plan + a. The present invention also relates to a sequence of nuclei + idos useful in the rne + odo. Rdernas, the present invention relates to a promoter * that is utL1 to express the core sequence + i two. Starch is one of the main storage carbohydrates in plan + as, especially higher plants. The structure of the starch consists of anulose and ironlopec + ina- The arnilose consists essential inen-t e of straight strands of linked glycosyl or-1-4 residues. The arnilopectma comprises chains of glyco residues ot-1-4 linked to some branches w-1-6. The branched nature of arn lopectin is achieved through the action of, in other things, an enzyme commonly known as the starch-breaking enzyme ("SBE"). The SEE catalyses the formation of branching points in the arn? Lopec + i na molecule by the addition of α-1,4 glucans through branching links or-1, 6-glucos? D? Cos. The biosintes of the anulose and the lopectin is shown schematic in Figure 1, while the bonds a-1-4 and the links a-l-ß are shown in figure?. It is known that starch is an important raw material. Starch is widely used in the vegetable, animal and paper industries. However, a large fraction of the starches that are used in these industrial applications is modified, after harvesting, by chemical, physical or other methods. enzyin + icos to obtain starches with certain functional properties required. In years r * ec? En + es, it has become desirable to obtain genetically modified plan + .as that may be capable of producing modified starches that may be * equal to modified post-harvest starches. It is also known that it may be possible to prepare such genetically modified plants by expressing nucleic acid coding sequences. In this regard, June Bourque provides a detailed summary of antisense strategies for genetic manipulations in plants (Bourque 1995, Plant Science 105 pp 125-149). When it is known that enzymatic activity can be affected by the expression of particular sequences of nucleotides (for example, see the teachings of Fmnegan and McElroy 1.1994] Biotechnology 12.883-888; and r1atzu.-e and riatzuke [1995] TTG Ll 1-3), there is still a need for a method that can affect enzymatic activity more reliably and / or more efficiently and / or specifically. In accordance with a first aspect of the present invention, a method is provided for affecting the enzymatic activity in a plant (or cell, tissue or organ thereof), which comprises expressing in the plant (or a cell, a tissue or an organ thereof) a nucleotide sequence, wherein the nucleotide sequence encodes, partially or completely, a m + ron in a sense orientation; and where the sequence of nucí eotí dos does not contain a sequence that is of sense, do a sequence of the exon normally associated with the m + ron. According to a second aspect of the present invention, a method is provided for affecting the enzymatic activity in a starch-producing organism (or a cell, a tissue or an organ thereof), which comprises expressing in the starch-producing organism. (or a cell, a tissue or an organ thereof) a sequence of nucleotides, wherein the nucleotide sequence encodes, partially or completely, an intron in a sense orientation; wherein the nucleotide sequence does not contain a sequence that is sense for an exon sequence normally associated with the intron; and wherein the activity of the starch branching enzyme is affected and / or apulopectin levels are affected and / or the composition of the starch is modified. According to a third aspect of the present invention, a sequence comprising the nucleotide sequence shown as any of T.D. is provided. SEC. No. 15 to I.D. SEC. No. 27, or a variant, derivative or homologous thereof.
According to a fourth aspect of the present invention, a promoter is provided which comprises the sequence shown as T.D. SEC. No. 14 or a variant, derivative or homologous thereof. In accordance with a fifth aspect of the present invention, a construct capable of understanding or expressing the present invention is provided. According to a sixth aspect of the present invention, a vector comprising or expressing the present invention is provided. According to a seventh aspect of the present invention, a cell, tissue or organ comprising or expressing the present invention is provided. According to an eighth aspect of the present invention, a transgenic starch producing organism comprising or expressing the present invention is provided. According to a ninth aspect of the present invention, a starch obtained from the present invention is provided. In accordance with a tenth aspect of the present invention, pBERB (NCIMB 40753) or pBEfig (NC1MB 40815) is provided. In accordance with the eleventh aspect of the present invention, a nucleotide sequence is provided which is antisense to any one or more of the sequence sequences obtained from? -SBE 3.2 (NCIMB 40751) or? - SBE 3.4 (NCIMB 40752) or a variant, derivative or homologous thereof. A key advantage of the present invention is that it provides a method for preparing modified starches and that it does not depend on the need to modify the postharvest starches, flsi, the method of the present invention avoids the need to use hazardous chemical compounds that are normally used in the modification of postharvest starches. Finally, the present invention provides, among other things, genetically modified plants which are capable of producing modified and / or novel and / or improved starches whose properties would satisfy various industrial requirements. The present invention provides a method for preparing adapted starches in plants that can replace modified postharvest starches. At the same time, the present invention provides a method for preparing modified starches by a method that may have an effect on or more beneficial to the environment than the known methods of postharvest modification, which depend on the use of compounds dangerous chemicals and large amounts of energy. Another key advantage of the present invention is that it provides a method that can affect the enzymatic activity more reliably and / or efficiently and / or more specifically in comparison with the methods known to affect the enzymatic activity. Regarding this advantage of the present invention, it should be noted that there is a close degree of homology between the coding regions of the SBEs. However, there is little or no homology with the intron sequences of the SBF-s. Physi, the anti-sitting expression of the mtron provides a mechanism that selectively affects the expression of a particular SBE. This advantageous aspect can be used, for example, to reduce or eliminate a particular enzyme of the SBF and replace this enzyme with another enzyme that can be another enzyme branched to, or even a recornbmante version of the affected enzyme, or a hybrid enzyme that can understand, for example, part of an SBE from one source and at least part of another SBE from another source. This particular character of the present invention is covered by the combination aspect of the present invention which is discussed in more detail below. In this case, the present invention provides a mechanism to selectively affect the activity of SBE. This contrasts with the methods of the prior art, which depend on the use of, for example, the anti-sense expression of the exon, according to which it would not be possible to introduce a new activity of the SBE without a ecta- also that acAvity. Preferably with the first aspect of the present invention, the activity of the starch enrichment enzyme is affected and / or where the levels of arnilopectma are affected and / or the composition of the starch is modified. Preferably with first or second aspect of the present invention, the nucleotide sequence does not contain a sequence that is sense to a sequence. Preferrably with the fourth aspect of the present invention, the promoter is in combination with an ether gene ("G01"). Preferably, the enzyme + ica activity is reduced or the? M? na Preferably, the nucleotide sequence l codes for * at least substantially at least? N intron in a sense orientation. Preferably, the nucleotide sequence encodes, partially or corn? Etamen + e, two or more introns, and where each intron is in a sense orientation. Preferably, the nucleotide sequence comprises at least 350 nucleotides (for example, at least 350 base pairs), more preferably at least 500 nucleotides (e.g., at least 500 base pairs). Prefei *? b1ernen + e, the sequence of nuc1 eot i two comprises the sequence shown as one of T, D. DE SEC. No. 1 to I.D. SEC. No. 13 or a variant, derivative or homologous thereof, including combinations thereof.
Preferably, the nucleotide sequence is expressed by a promoter having a sequence shown co or I.D. SEC. No. 14 or a variant, derivative? homologo of the same. Preferably, the transgenic organism that produces the starch is a plant. A preferred aspect of the present invention therefore relates to a method for affecting the enzymatic activity in a plant (or a cell, a tissue or an organ of the same), which comprises expressing * in the plant (or a cell, a tissue or an organ thereof) a sequence of nucleotides, wherein the nucleotide sequence encodes, partially or completely, a mtron in a sense orientation; wherein the sequence of nuci eotides does not contain a sequence that is sense to an exon sequence normally associated with the mtron; and wherein the activity of the starch enrichment enzyme is affected and / or the levels of arnilopectma are affected and / or the starch composition is modified. A more preferred aspect of the present invention therefore relates to a method for affecting the enzymatic activity in a plant (or a cell, a tissue or an organ thereof), which it is necessary to express in the plant ( or a cell, a tissue or an organ thereof) a sequence of nucleotides, wherein the sequence of n? c eotides encodes, partially or completely, a mtron in a sense orientation; wherein the nucleotide sequence does not contain a sequence that is sense to an exon sequence normally associated with the mtron; wherein the activity of the starch branching enzyme is affected and / or the levels of arylopectin are affected and / or the starch composition is modified; and wherein the nucleotide sequence comprises the sequence shown as any of T.D. OF SFC. No. 15 a
I.D. SEC. No. 27, or a variant, depvado or homologo of the same, including combinations of the same. The term "nucleotide", in relation to the present invention, includes RDN and flRN. Preferably it means RDN, more preferably RDN prepared by the use of recornbinating RDN techniques. The term "intron" is used, in its normal sense, to denote * a segment of nucleotides, often RDN, that does not encode an entire expressed protein or enzyme, or part thereof. The term "exon" is used, in its normal sense, to indicate a segment of nucleotides, often RDN, that encodes a whole expressed protein or enzyme, or part of the miRNA. In addition, the term "ntron" refers to regions of genes that are transcribed into NRN molecules, but that bind outside the NRN before the RRN is translated into a protein. In contrast, the term "exon" does not refer to genes that are transcribed in RRN and that are subsequently translated into proteins. The terms "variant" or "homologous" or "fragment" in relation to the nucleotide sequence of the present invention, include any substitution of, variation of, modification of, replacement of, deletion (loss) of, or addition of, one (or more) nucleic acids for the respective n-cleotide sequence, provided that the resulting nucleotide sequence + can affect the onzirnatic activity in a plant, or cell or tissue thereof, preferably wherein the resulting nucleotide sequence has at least the same effect as any of the sequences of sense shown as TD SEC. Nos. 1-13. In particular *, the term "homologous" covers the homology with respect to the similarity of structure and / or similarity of function, provided that the resulting nucleus sequence + idos has the ability to affect the enzymatic activity in accordance with the present invention. . In relation to the homology (i.e., similarity) of the sequence, I strongly preferred there to be 80% homology, more preferably at least 85% homology, more preferably at least 90% homology, even more preferably at least minus 95% homology, but I preferred blernente at least 98% homology. The above terms are also synonymous with allelic variations of the sequences. The terms "variant" or "homologous" or "fragment" in relation to the nucleotide sequence of the present invention, include any substitution tle, variation of, modification of, replacement of, deletion (loss) of, or addition of, one (or more) nucleic acids for the respective nucleotide sequence, provided that the resulting promoter sequence permits the expression of a GOI, preferably wherein the resulting nucleotide sequence has at least the same effec + or (th) SECTION 14. SECTION 14. In particular, the term "homologous" covers homology with respect to the similarity of structure and / or similarity of function, provided that the resulting nucleotide sequence has the capacity to allow the expression of a GOI, such as a nucleotide sequence according to the present invention With regard to the homology (ie, *, similarity) of the sequence, preferably there is more than 80% homology, preferably at least 85% homology, preferably by at least 90% homology, even more preferably at least 95% homology, preferably at least 98% homology. The above terms are also synonymous with allele variations of the sequences. The sequence of introns of the present invention can be any or all of the sequences of the voyeurs of the present invention, including partial sequences thereof, provided that, if sense sequences are used (i.e., sequences that do not comprise some or more of the complete sequences shown as SEQ ID No. 1-13), the partial sequences affect the enzymatic activity. Suitable examples of partial sequences include sequences that are more correct than any of the complete sense sequences shown as TD. of SEC. No. 1-L3 but comprising nucleotides that are adjacent to the exon or exons thereof. With respect to a second aspect of the present invention (ie, specifically affecting the activity of SBE), the nucleoside sequences of the present invention may comprise one or more sequences of sense or antisense exons or of the SBE gene (but there are no sequences of sense exons naturally associated with the sequence of introns), including complete or partial sequences thereof, provided that the nucleotide sequences can affect SBE activity, preferably where the nucleotide sequences reduce or eliminate the activity of SBE. Preferably, the nucleotide sequence of the second aspect of the present invention does not comprise sequences of sense exons. The term "vector" includes an expression vector and a transformation vector. The term "expression vector" means a construct capable of m-vivo or m-vitro expression. The term "transformation vector" means a construct capable of being * transferred from one species to another, such as from a plasmid of E. coli to a fungus or a plant cell, or from flrobactep to a plant cell. The term "construct", which is synonymous with such terms as "conjugate", "cassette" and "hybrid", in relation to the antisense nucleotide sequence aspect of the present invention, includes the nucleotide sequence of conformance. with the present invention linked directly or indirectly to a promoter. An example of an indirect link is the provision of an appropriate separating group * such as a sequence of the pattern, such as the? Ntr * on Shl or the nt ron RDH, int medium to the promoter and the nucleotide sequence of the present invention. The same is true for the term "merged" in relation to the present invention, which includes direct or indirect linkage. The terms do not cover the natural combination of the SBE gene of the wild type when it is associated with the wild-type SBE gene promoter in its natural environment. The construct can contain or even express a marker that allows the selection of the direct genetic construct, for example, in a plant cell in which it has been transferred. There are several markers that can be used, for example, in plants - such as handy. Other examples of markers include those that provide resistance to antibiotics - for example, resistance to G418, hl romicin, bleomicma, canarnicma and gentarnicin. The construct of the present invention preferably comprises a promoter. The term "promoter" is used in the normal sense of the art, for example, a binding site for RRN in the Jacob and Monod theory of gene expression. Examples of suitable promoters are those that can direct the efficient expression of the nucleotide sequence of the present invention and / or in a specific type of cell. Some examples of tissue-specific promoters are described in wO 92/11375. The promoter may include * additionally from conserved regions such as a Pprnbow block or a TflTR block. The promoters may contain even other sequences that affect (so that they maintain, increase, decrease) the levels of the Lon Express of the nucleotide sequence of the present invention. Suitable examples of said sequences include the intron Shl or an intron RDM. Other sequences include mobile elements - such as temperature, chemical agents, light or stress-causing elements. Also, suitable elements that increase transcription or translation may be present. An example of the last element is the main sequence TflV 5 '(see Sleat Gene 217 Cl 987-1 217-225; and Dawson Plan + Mol. Biol. 23 C1993] 97). As mentioned above, the construct and / or vector * of the present invention can include a region of transcriptional axis that can provide regulated or constitutive expression. Any suitable promoter can be used for the region of the start of transcription, such as a tissue-specific promoter. In one aspect, preferably the promoter is either a palatin promoter or the E35S promoter. In another aspect, preferably the promoter is the SBE promoter. If, for example, the organism is a plant, then the promoter may be one that affects the expression of the nucleotide sequence in any one or more tissues of the seed, tuber, stem, shoot, root and leaf. Preferably + tuber. As an example, the promoter-for the nucleotide sequence of the present invention may be the a-Arny 1 promoter (also known as the Rmy 1 promoter, the Amy 637 promoter or the 'cc-Arny promoter (537) as described in UK co-pending patent application No. 9421292.5 filed on October 21, 1994. Alternatively, the promoter for the nucleotide sequence of the present invention may be the a-Arny 3 promoter (also known as the Rrny promoter). 3, the promoter * Rmy 351 or the promoter a-Riny 351) as described in co-pending UK patent application No. 9421286.7 filed on October 21, 1994. The present invention also encompasses the use of a promoter that expresses a sequence of nucleotides according to the present invention, wherein a part of the promoter is mac + ivated, but where the + orymbor may still function as a promoter.Partial inactivation of the promoter is in some cases, advantageous. In particular, in the case of the promoter * Rrny 351 mentioned above, it is possible to inactivate a part thereof in which the partially inactivated promoter expresses the nucleotide sequence of the present invention in a more specific manner such as in a type of tissue or specific organ. The term "inactivated" means partial inactivation in the sense that the paf fon of expression of the promoter is modified, but where the IB
partial promoter + e activated works even as a promoter. However, as mentioned above, the promoter is modified capable of expressing a gene encoding the enzyme of the present invention in at least one specific tissue (but not all) of the original promoter. Examples of partial inactivation include altering * the folding pattern of the promoter sequence, or joining species to parts of the nucleotide sequence, so that a part of the nucleotide sequence, not recognized by, eg, the RRN polymerase Another (preferable) way to partially inactivate the promoter is to truncate it to form fragments thereof. Another way would be to mule at least a part of the sequence so that the polynomial RRN can not join that pair + e or another part. Another modification is rnu + ar the si + ios the union for regulatory proteins as, for example, the protein Crefl ele filainen fungi + bears that are known to repress the catabolite by * carbon, and thus suppresses the repression ejel catabolito of the promoter * native . The construct and / or vector of the present invention may include a region of transcription terms. The nucleotide according to the present invention can be expressed in combination (but not necessarily at the same time) with an additional construct. Rsi, the present invention also provides a combination of constructs comprising a first construct comprising the nucleotide sequence according to the present invention operably linked to a first promoter *; and a second construct comprising a GOT operatively linked to a second promoter (which need not be the same as the pprner promoter). With this aspect of the present invention, the combination of constructs can be present in the same vector, plasmid, cells, tissue, organ or organism. This aspect of the present invention also covers methods of expression thereof, preferably in specific cells or tissues, such as expression in a cell or tissue specific to an organism, typically a plant. With this aspect as the present invention, the second construct does not cover the natural combination of the gene encoded for an enzyme ordinarily associated with the wild-type gene promoter when both are in their natural environment. An example of a suitable combination would be a first construct comprising the nucleotide sequence of the present invention and a promoter, such as the promoter of the present invention, and the second construct comprising a promoter *, such as the promoter of the present invention. invention, and a GOI in which the GOI codes for another starch branching enzyme either in sense or antigenic orientation. The above comments relating to the term "construct" for the antisense nucleotide aspect of the present invention are the same as applicable to the term "construct" for the promoter aspect of the present invention, in this respect, the term includes the promoter according to the present invention directly or indirectly bound to GOL The term "GOI" with reference to the promoter aspect of the present invention or to the combination or aspec + of the present invention means any element gene, which does not need a code for a pro + eine or an enzyme - as explained later. A GOI can be any nucleus sequence + idos that is foreign or natural to the organism, in question, for example, a plant. Typical examples of a GOI include genes that code for other proteins or enzymes that modify methicine and catabolic processes. GOT can code for an agent to introduce or increase resistance to pathogens. The GOI can even be an antisense construct pair * to modify the expression of natural transcripts present in the relevant tissues. An example of said GOI is the nucleotide sequence according to the present invention. The GOI can even code for a protein that is non-natural for the host organism and / or, for example, a plant. The GOI can code for a compound that is of benefit to animals or humans. For example, GOI could code for a pharmaceutically active protein or enzyme such as any of the therapeutic compounds insulin, interferon, human serum alumina, human growth factor *, and blood agglutination factors. GOT can even code for a protein that gives additional citrus nutp value to a food or feed for livestock or crop. Typical examples include plant proteins which can inh the formation of antimatter factors and plant proteins having a desirable amino acid composition (e.g., a higher lysine content than a non-transgenic plant). AND? 1 GOI can even code * for an enzyme that can be used in food processing such as xylanases and w-galactosidase. The GOT may be a gene that codes for either a toxin from a pest, an antisense transcript such as or-arnilasa, a protease or a glucanase. Alternatively, the GOI may be a sequence of methods according to the present invention. G01 may be a sequence of nucleotides encoded for the arabinofuranosidase enzyme which is the subject of our co-pending United Kingdom Patent Application No. 9505479.7. The GOI may be the sequence of nucleotides, eg it encodes the enzyme gluca asa, which is the triple of the co-pending U.S. Patent Application No. 9505475.5 of the present inventors. The GOT may be * the sequence of nucleotides encoding the enzyme a-arnilasa which is the triple of co-pending U.S. Patent Application No. 9413439.2 of the present inventors. The GOT may be the nucleotide sequence that codes for the enzyme a-arnilasse, which is the subject of copending United Kingdom Patent Application No. 9421290.9 of the present inventors. The GO can be any of the nucleotide sequences encoding the a-gl? Canl enzyme that are described in copending PCT Patent Application No. PCI / EP94 / 03397 of the inventors herein. In one aspect, the GOI may even be a sequence of nucleotides according to the present invention but when it is operably linked to a different promoter. The GOT could include a sequence that codes for one or more than one xylanase, one arabic loop, one acetyl esterase, one rarnnogalactinase, one glucanase, one pect mass, one branching enzyme or another carbohydrate or protein modifying enzyme. Alternatively, the GOI may be a sequence that is antisense to any of those sequences. As mentioned before, the present invention provides a mechanism to selectively affect a particular enzymatic activity. In an important application of the present invention, it is now possible to reduce or eliminate the expression of a genomic nucleotide sequence encoding a protein or genomic enzyme by expressing an antisense intron construct for that particular protein or gene enzyme and ( for example at the same time) expressing a recornbmante version of that enzyme or pro-teí to - in o + ras words, the GOT is a sequence of nucleus + recon ante es that codifies for the enzyme or genomic protein. The expression of enzymes and recombinant proteins desired in the absence (or reduced levels) of respective enzymes and gene proteins is + to application perrni + e. Therefore, the desired recombinant enzymes and pro-emines can be easily separated and purified from the host organism. This particular aspect of the present invention is very advantageous over prior art methods * which, for example, are based on the use of antisen + exon expression, said methods also affect the expression of the recornbinating enzyme. Therefore, a further aspect of the present invention relates to a method of expressing a recombinant protein or enzyme in a host organism which comprises expressing a sequence of nucleotides encoding the recombinant enzyme or protein.; and that said a sequence of nucleotides in which the nucleotide sequence encodes, partially or completely, for a m + rn in an anti-sense orientation; wherein the intron is a mtron normally associated with the genomic gene encoding a protein or an enzyme corresponding to the protein or enzyme recornbinan + e; and wherein the additional nucleotide sequence does not contain a sequence that is antisense to an exon sequence normally associated with the ín + ron. Additional aspects cover the combination of the nucleotide sequences including their incorporation into constructs, vectors, cells, tissues and transgenic organisms. Therefore, the present invention also relates to a combination of nucleotide sequences comprising a first nucleotide sequence which codes for a recombinant enzyme; and a second nucleotide sequence corresponding to a tron in antisense orientation; wherein the mtron is an intron, which is associated with the genoinic gene that encodes an enzyme corresponding to the recornbinating enzyme; and wherein the second nucleotide sequence does not contain a sequence that is antisen + gone for an exon sequence normally associated with the intron. The GOT may even code for one or more rodents, such as any one or more of the intron sequences presented in the attached sequence lists. For example, the present invention also covers the expression, of, for example, an an + isentide mtron (eg, SEQ ID No. 15) in combination, for example, with an eletronic monomer. which preferably is not complementary to the intron sequence of antLsen + ido (e.g., TD of SEC.2). The term "cell", "tissue" and "organ" includes cell, tissue and organ per se and when within an orgaru srno. The term "organism" in connection with the present invention includes any organism that can comprise the nucleotide sequence according to the present invention and / or wherein the nucleotide sequence according to the present invention can be expressed when present in the organism. Preferably, the organism is an organism such as starch products or any of a plant, algae, fungus, yeast and bacteria, as well as cell lines thereof. Preferably, the organism is a plant. The term "starch-producing organism" includes any organism that can biosynthetize starch. Preferably, the starch producing organism is a plant. The term "plant", as used herein, includes any angiosperm, gi nosperm, monocotyledonous and dicotyledonous. Typical examples of suitable plants include vegetables such as potatoes; cereals such as wheat, rnaíz and barley; fruits; Trees flowers and other plant crops. Preferably, the term means "potato." The term "transgenic organism" in relation to the present invention includes any organism comprising the nucleotide sequence according to the present invention and / or products obtained therefrom, and / or wherein the nucleotide sequence according to the present invention invention can be expressed * within the organism. Preferably, the nucleotide sequence of the present invention is incorporated into the genome of the organism. Preferably, the transgenic organism is a plant, preferably preferably a potato. For preparing the host organism prokaryotic or eukaryotic organisms can be used. Examples of suitable procapotide hosts include E. coli and Bacillus sub + ilis. The teachings on the transformation of procapotic hosts are well documented in the art, for example, see Sarnbrook et al. (Sarnbroo et al. In Molecular Clone g: A Laboratory Manual, 2nd edition, 1989, Cold Spring Harbor-Laboratory Press). Although the enzyme according to the present invention and the nucleus sequence + encoding thereto are not described in EP-B-0470145 and CR-R-200645, these two documents provide useful supporting comments on the + + techniques that can be employed to prepare transgenic plants in accordance with the present invention. Some of these supporting teachings are now included in the following commentary. The basic principle in the construction of plan + as genet? Carne + e modified is mser + ar genetic information in the genorn of the plant to obtain a stable maintenance of the genetic material inserted. There are several - techniques for inserting * genetic information, the two most important principles being the direct introduction of genetic information and the introduction of genetic information through the use of a vector system. A review of the general techniques can be found in articles by Potry.-us (Annu Rev Plan + Physiol Plant Mol Biol T1991] 42: 205-225) and Chpsto (Agro-food-Industry Hi-Tech March / Appl 1994 1 7- 27). Therefore, in one aspect, the present invention relates to a vector system that carries a sequence of nucleotides or construct in accordance with the present invention and that is capable of introducing the nucleotide sequence or construct into the geno of an organism, such as a plant. The vector system may comprise a vector, but may also comprise two vectors. In the case of two vectors, the vector system is usually referred to as a binary vector system. The binary vector systems are described more fully in Gynheung An et al., (1980), bmary Vec-tors, Plant Molecular Biology Manual A3, 1-19. A system widely used for the transformation of plant cells with a + or nucleus sequence or a given construct is based on the use of a Ti plasmid from figrobactermrn tu efaciens or a Ri plasmid from Agrobacterium rhizogenes Pn and others (1986),
Plant Physiol, 81, 301-305 and B? Tcher D.N. and others (1980), Tissue Culture Methods for Plant Pathologists, eds .: D.S. Ingrarns and J.P. Helgeson, 203-208. Several different Ti and Ri plasmids have been constructed which are suitable for the construction of previously described plant or plant cell constructs. A non-limiting example of said 2b
Plasmid Ti is pGV3850. A nucleotide sequence or spindle construct the present invention should preferably be inserted into the Ti plasmid between the terminal sequences of the T-DNA or a T-DNA sequence to avoid * the alteration of the sequences immediately surrounding the boundaries of the T-DNA , at least one of these regions appears to be essential for the insertion of T-DNA into the plant genotype. As understood from the above explanation, if the organism is a plant, the vector system of the present invention is preferably one that contains the sequences necessary to infect the plant (e.g., the region). n vir) and at least a borderline part of a T-DNA sequence, the borderline part being located on the same vector as the genetic construct. In addition, the vector system * is preferably a Ti axis Agro ba c t e r i uin t r ume f a c i e n s a plasmid Ri ele
Agrobactopurn rhizogenes or a derivative thereof. For these plasmids are well known and widely used in the construction of trangerucas plants, there are many vector systems that are based on these plasmids or derivatives thereof. In the construction of a transgenic plant, the nucleotide sequence or contruct of the present invention can first be constructed in a microorganism in which the vector can be replicated and which is easy to manipulate before inserting into the plant. An example of a u + il organism is E. coli, but other organisms having the above properties can be used. When a vector or a vector-like system was defined as an + it has been constructed in E. coli, it is transferred, if necessary, into a suitable strain.
Agrobac + enurn, e.g., Pgrobact e ri urn + urne f aciens. The plasmid Ti carrying the nucleotide sequence or cons + ruct of the present invention is thus preferably transferred to a suitable Agrobactepurn species eg, A. turnefacies, to obtain an Agrobacterium cell which carries the promoter or nucleotide sequence or construct of the present invention, said DNA being subsequently tansphobic towards the cell of the plant to be * modified. If, for example, for the transformation the Ti or Ri plasmid of the plant cells is used, at least the right border and often the right border and the left border of the T-RDN of the plasinid Ti and the plaspudo Ri, as Flanking areas of the introduced genes can be connected. The use of T-RDN for the transformation of plant cells has been studied intensively and is described in EP-fl-120516; Hoe ema in: The Binary Plant Vector System Offset-drukl - erij Kanters B.B., Rlblasserdarn, 1985, Chapter V; Fraley et al., Cpt. Rev. Plant Sci., R: l-4B; and fln et al., EMBO J. (1985) 4: 277-284. The direct infection of plant tissues by flgrobacteriurn is a simple technique that has been widely used and described in Butcher D.N. and others (1980), Tissue Culture Methods for Plant Pathologists, eds .: D.S. Tngrarns and J.P. Helgeson, 203-208. For additional teachings on this subject, see Potry us (Annu Rev Plant Physiol Plant MolBiol [1991] 42: 205-225) and Chpsto? (Agro-Food-Tnelust r and HiTech March / April 1994 17-27). With this technique, the infection of a plant can be done inside or on a certain part or tissue of the plant, that is, on a part of a leaf, a root, a stem or another part of the plant. Typically, with direct infection of plant tissues *, by Agrobactepum carrying the GOI (such as the nucleotide sequence according to the present invention) and optionally a promoter *, a plant that is going to be infected is injured, eg, cutting the plant with a blade of shaving or pricking the plant with a needle or rubbing the plan + a with an abrasive. The lesion is then inoculated with Agrobac + epurn. The plan + a or par + e of the inoculated plan + is then grown on a suitable culture medium to allow development in mature plants. When plant cells are constructed, these cells can be cultured and maintained according to well-known tissue culture methods such as by culturing the cells in a suitable culture medium supplied with the necessary growth factors such as amino acids, plant hormones. , vitamin, etc. The regeneration of transformed cells in genetically modified plants can be achieved using known methods for the regeneration of plants from cell or tissue cultures, for example, by selecting transformed offspring using an antibiotic and subculturing the rods in a medium containing the appropriate nutrients, vegetole hormones, etc. Additional teachings about the transformation of plants can be found in EP-A 0449375. As reported in CA-A-2006454, a large number of cloning vectors containing a replication system in E is available. co l i and a marker that allows a selection of the transformed cells. The vectors contain by * example pBR 322, pUC series, M13 series rnp, pRCYC 184 etc. In this manner, the nucleotide or construct of the present invention can be introduced into a suitable restriction position in the vector *. The content plasmid is then used for transformation into E. coll. The E. coli cells are grown in an appropriate nutrient medium and then harvested and lysed. The plasmid is then recovered. A sequence analysis, restriction analysis, electrophoresis and biochemical and molecular biology methods are generally used with a method of analysis. After each manipulation, the RDN sequence used can be restricted and connected to the next DNA sequence. Each sequence can be cloned in the plasmid state or in a different one. After the introduction of the sequence of nucleotides or contructo axis compliance with the present invention in the plants, the presence and / or insertion of additional RDN sequence may be necessary - so as to create combination systems as I indicated before (for example, an organism that comprises a combination of constructs). The above commentary for the transformation of prokaryotic organisms and plants with the sequence of nuclei of the present invention is equally applicable to the transformation of those organisms with the + orifice of the present invention. In summary, the present invention relates to affecting the enzymatic activity by expressing antigenic sequences of the antigen. Also, the present invention relates to a promoter useful for the expression of those sequences of the tron of anta sense. The following samples have been deposited according to the Treaty of Budapes + in the recognized deposit The National Colloctions of Industrial and Marine Bacteria Limited
(NCTMB) to 23 St Machar Dpve, flberdeen, Scotland, AB2 1RY, Rowing
Joined, July 13, 1995: NCIMB 40754 (which refers to pBEA 11 as described herein); NCIMB 40751 (which refers to? -SBF 3.2 as described here), -and CIMB 40752 (which refers to? -SBE 3.4 as described here). A highly preferred embodiment of the present invention, therefore, relates to a method for affecting the enzymatic activity in a plan + a (or a cell, or a + e? Do or an organ of the same) which comprises expressing in the plant (or a cell, or a tissue or an organ of the same) a sequence of nucleotides in which the nucleotide sequence encodes partially or completely for? n intron in a sense orientation; wherein the nucleotide sequence does not contain a sequence that is sense for an exon sequence normally associated with the mtron; in which the enzymatic activity of starch branching is affected and / or the levels of arm lopectma are affected and / or the starch composition is changed; and wherein the nucleotide sequence can be obtained from NCIMB 40751, NCTMB 40752 or NCTMB 40754, or a variant, derivative or homologous thereof. A highly preferred aspect of the present invention, therefore, relates to a method for affecting enzymatic acivity in a plan + a (or a cell, a tissue or an organ thereof) comprising expressing * in the plant (or a cell, a + e? or an organ thereof) a sequence of nucleotides in which the nucleotide sequence encodes, partial or corn? le + amenté, for an in + ron in a sense orientation; where the sequence of nucleo + idos not with + iene a sequence that is of sense for a sequence of normal exon men + e associated with the in + ron; wherein the activity of the starch branching enzyme is affected and / or the levels of the arm lopecti na are affected and / or the starch composition is changed; wherein the sequence of nuclei + idos comprises the sequence shown as any of SEQ ID No. 1 to SEQ ID No. 13 or a variant; derivative or homologous of the ism, including combinations thereof; and wherein the nucleotide sequence is obtainable from NCIMB 40751, NCTMB 40752 or NCTMB 40754, or a variant, derivative or homologous thereof. The present invention will now be described by way of example, in which reference is made to the following appended figures: Figure 1 is a schematic representation of the biosynthesis of anulose and arnilopectma; Figure 2 is a diagrammatic representation of the a-1-4 bonds and the a-l-6 bonds of arnilopema; Figure 3 is a diagrammatic representation of the exon-mt on structure of a genomic SBE clone; Figure 4 is a plasmid map of pPATAl, which is 3936 base pairs in size; Figure 5 is a map of axis plasmid? ABE7, which is 5106 base pairs in size; Figure 6 is a plasmid map of pVictorlV Man, which is 7080 base pairs in size; Figure 7 is a plasmid map of pBEAll, which is 9.54 kb in size;
Figure 8 shows the complete genomic nucleotide sequence for SBE including the promoter, exons and estrogens; Figure 9 is a plasmid map of pVictorda, which is 9.1? kb in size; and Figure 10 is a plasma map of pBEP2, which is 10.32 kb in size; Figures 1 and 2 were mentioned above in the introductory description referring to starch in general. As mentioned, Figure 3 is a diagrammatic representation of the exon-mtron structure of a genomic SBE clone, whose sequence is shown in Figure 8. This clone, which is approximately 11.5 kb, comprises 14 exons and 13 int roñes. The patterns are numbered in increasing order from the 5 'end to the 3' end and correspond to SEQ ID Nos. 1-13, respectively. Their respective ant i sense intron sequences are shown as SEC ID No. 15-27. With more detail, the Figures 3 and 8 present information about 11468 base pairs of the potato SBE gene. The 5 'region of nucleotides 1 to 2082 contains the promoter region of the SBE gene. A TATA box candidate in nucleotide 2048 through 2051 is locked in a ca. The homology between a SBE cDNA clone of potato (Poulsen S Kreiberg (1993) Plant Physiol 102: 1053-1054) and the DNAs of exons being at 2083 bp and ending at 9666 bp. The homology between the cDNA and the DNA of the exons is indicated by the nucleofides with uppercase letters, while the translated sequences of the amino acids in the code are shown with unique letters under the DNA of the exons. The sequence of the letters is indicated with lowercase letters. Figure 7 is a plasinid map of pBER7, which is 9.54 kb in size. The plasmid pBER 11 bought the first SEB sequence of the potato. The first intron sequence, which has 1177 base pairs, is shown in Figure 3 and is between the first * exon and the second exon. These experiments and aspects of the present invention are now considered in more detail.
EXPERIMENTAL PROTOCOL
ISOLATION, SUBCLONRCION IN PLASMIC, AND SEQUENCING OF GENETIC SBE CLONES
Several clones containing the potato SBE gene were isolated from a genomic library of potato Desiree (Clontech Laboratories Inc., Palo Alto CA, USA) using radiolabelled potato SBE cDNA as a probe (Poulsen &Kreíberg ( 1993) Plant Physiol. 102: 1053-1054). The fragments of the isolated? -fagos containing SBE DNA (? SBE 3.2-NCTMB 40751- and? SBE-3.4-NCIMB 40752) were identified by * Southern analysis and then subcloned into the pBluescppt II vectors (Clontech Laboratories Inc. , Palo Rifo Cfl, USA.,? SBE 3.2 contains a potato RDN insert of 15 kb and? SBE-3.4 contains a potato RDN insert of 13 kb.The resulting plasmids were called? GB3, pGBll, pGB15, pGBld and? GB25 (see further discussion) The respective inserts were sequenced using the Pharmacia Autoread Sequencmg kit (Pharmacia, Uppsala) and an ALF DNA sequencer * (Pharmacia, Uppsala) In total, a stretch of 11.5 was sequenced. kb of the SBE gene We derived the sequence of the aforementioned amphibian plasmids, in which:? GB25 contains the sequences from 1 bp to 836 bp, pGB15 contains the sequences from 735 bp to 2580 bp,? GB16 cont lene The sequences ele 2580 at 5093 bp, pGBll contains the sequences from 3348 bp to 7975 bp and pGB3 with It has sequences from 7533 bp to 11468 bp. In more detail, a? GB3 was constructed by inserting a 4 kb EcoRI fragment isolated from? SBE 3.2 to the EcoRI site of pBluescppt II SK (+). A pGBll was constructed by an Xhol fragment of 4.7 kb isolated from > .SBE 3.4 to the Xhol site of pBlueecppt II SK (+). A? GB15 was constructed by a 1.7 kb Spel fragment isolated from / \ SBE 3.4 to the Spel site of pBluescnpt II SK (+). PGBlb was constructed by insertion of a 2.5 kb Spel fragment isolated from? SBE 3.4 to the Spel site of pBluescnpt II SK (+). For the construction of pGB25 a PCR fragment was produced with the primers 5 'GGA ATT CCA GTC GCA GTC TAC ATT AC 3'
'CGG GAT CCR GAG GCR TTA AGfl TTT CTG G 3
and \ SBE 3.4 as a template. A PCR fragment was digested with BarnHI and EcoRT, and inserted into pBluesc ipt II SK (+) digested by the same restriction enzymes.
CONSTRUCTION OF PLASMIDE pBEAll
The SBE intron 1 was amplified by PCR using the oligonucleotides:
'CGG GRT CCA AAG AAA TTC TCG AGG TTR CAT GG 3' and 5 'CGG GRT CCG GGG TRR TTT TTA CTA ATT TCA TG 3'
and the SBE 3.4 phage containing the SBE gene as template. The PCR product was digested with BarnHI and inserted in a sense orientation into the BarnHT site of the plasmid pPATAI (described in UO 94/24292) between the patatma promoter and the 35S ter-driver. This construction was digested? ABE7 with Kpnl, and the Kpnl fragment of "promoter axis patati na-? Ntr * on 1 of SBE-terrnmador 35S" of 2.4. b was isolated and inserted into the site
Kpnl of the plant transformation vector pVictorlV Man producing the plasinid pBEAll.
PRODUCTION OF TRANSGENIC PLANTS OF POTATO
Axénica Class Crops Cultivation of Solanum tuberosum 'Bintje' and 'Dianella' shoots on a substrate (LS) of a formula according to Linsrnaier, E.U. and Skoog, F. (1965), Physiol. Plan +. 18: 100-127, which also contains 2 μM of thiosulfate + or silver at 25 ° C and 16 hours of light / 8 hours of darkness. The cells were subcultured after approximately 40 days. The leaves were then cor + oros and cut into segmented nodes (approximately 0.8 crn) each containing a node.
Inoculation of Potato Tissues Crop stems were cut with rods of approximately 40 days (approximate height of 5-6 c) internodal instruments (approximately 0.8 crn). The segments were placed on liquid LS substrate containing transformed Rgrobacterium t? Mefaciens containing the binary vector of antibodies. Rgrobacteri urn was grown overnight in substratum YMB (dipotassium acid phosphate, tphidratatado (0.66 g / 1); magnesium sulfate, heptahydrate (0.20 g / 1); sodium chloride-uro (0.10 g / 1); rnamtol (10.0 g / 1); and yeast extract (0.40 g / D) containing appropriate antibiotics (corresponding to the resistance gene of the Agrobacterium strain) at an optical density at 660 nrn (OD-660) of approximately 0.8, centrifuged and resuspended in the LS substrate to an OD-660 of 0.5. The Agrobacten ™ rn suspension segments were left for 30 minutes and then the excess bacteria were removed by staining the segments on sterile filter paper *.
Co-culture The segments of the rod were co-cultured with bacteria for 8 hours directly on the LS substrate containing acid 2, -d? chlorofenoxiacet ico (2.0? ng / l) and trans-zeat a (0.5 rng / 1). The substrate and also the explants were covered with filter papers and the Petp boxes were placed at 25 ° C and 16 hours light / 8 dark.
"Washing" Procedure After 48 hours on the coculture substrate, the segments were transferred to vessels containing liquid LS substrate containing 800 mg / l carbenicillin. The containers were shaken gently and by this procedure either they were separated by washing the segments and / or most of the Agrobacterium was killed.
Selection After the washing procedure, the samples were transferred to plates containing the IS substrate, agar * (8 g / 1), trans-zeatin (1-5 mg / l), gibberellic acid (Ol rng / 1), carbemcalin (800 mg / l) and canamic sulfate (50-100 rng / 1) or phosphonated na (1-5 rng / 1) or mannose (5 g / 1) depending on the construction of the vector used. The segments are subcultured to fresh substrate every 3A weeks. In 3 to 4 weeks, the rods were developed from the segments and the formation of new rods continues for 3-4 months.
Rooting of the Regenerated Stems V The regenerated rods were transferred to rooting substrates composed of LS substrate, agar (8 g / 1) and carbenicilm (800 rng / 1). The transgenic genotype of the regenerated stem was verified by examining the rooting capacity on the aforementioned substrate containing canarnicma sulphate (200 rng / 1), performing the NPTII S.E. and others, Theor. Appl. Genet (1988), 7: 68-694) and performing the PCR analysis according to Wang et al. (1993, NAR 21, pp4153-4154). Plants that were not positive in these trials were discarded or used as controls. Alternatively, the transgenic plants could be verified by performing a GUS assay on the β-glucuronidase gene or introduced according to Hodal, L. et al. (Pl.Sci. (1992), 87: 11-122).
Transfer to Soil Freshly rooted plants (approximate height of 2-3 crn) of the rooting substrate were transplanted to the soil and placed in a culture chamber (21 ° C, 16 hours light 200-400uF / rn2 eg). When the plants were well established, they were transferred to the greenhouse, where they were grown until the tubers had developed and the upper part of the plants was aging.
Harvest The potatoes were harvested after approximately 3 months and then analyzed.
ANALYSIS OF THE BRANCH ENZYMES
SBE expression was determined in transgenic potato lines using the SBE assays described by Blennow and Johansson (Phytochemí st r * y (1991) 30437-444) and by normal Uestern procedures using antibodies directed against potato SBE.
STARCH ANALYSIS
Starch was isolated from potato tubers and smoked in terms of the ratio: ami lopecti na (Hovenkarnp-Herrnelmk et al. (1988) Potato Research 31: 241-246). The distribution of the chain length of the ilopectin was determined by analyzing the starch digested by the isoamylase on a Dionex HPAEC. The number of reducing ends of the starch digested by isoarnilase was determined by the method described by N. Nelson (1994) J. Biol. Chern. 13: 37-380. The results revoked that there was a reduction in the level of synthesis of the SBE and / or the activity level of the SBE and / or the composition of the SBE of the starch in the transgenic plants.
TRAINING OF THE CONSTRUCTQ PROMOTOR of SBE
A promoter fragment axis SBE axis amine? ~ SBE 3.4 was used using the primers:
'CCR TCG RTR CTT Tflfl GTG ATT TGA TGG C 3'
'CGG GAT CCT GTT CTG ATT CTT GAT TTC C 3'
The PCR product was digested with Clal and BamHl. The resulting 1.2 kb fragment was then inserted into pVictorda (see Figure 9) linearized with Clal and Bg /? ? producing PBEP2 (see Figure 10).
MEASUREMENTS OF THE STARCH BRANCH ENZYMES OF POTATO TUBERS
Potatoes were cut from the potato plants transected with either pBEAll in small pieces and they were homogenized in extraction regulators (0 rnM Tps-HCI pH 7., Sodium dationite (0.1 g / 1) and 2 mM DTT ) Ultra-Turax; 1 g of Dowex xl was added per 10 g of tuber. The crude homogenate was filtered through a mirate filter and centrifuged at 4 ° C for 10 minutes at 24-700 g. A supernatant was used for the testing of the starch enrichment enzyme. The starch enzyme assays were carried out at 25 ° C in a volume of 400 μl composed of 0.1 μM of regular pH 7.0, of sodium treatment, 0.75 μg / μl of anulose, 5 μg / μl of bovine serum albumin and potato extract. At 0, 1, 30 and 60 minutes the aliquots of 50 μl to 20 μl of 3 N HCl were removed from the reaction. 1 nrn in iodine solution was added and the decrease in absorbance was measured at 620 nrn with a rothotorne specimen or ELISA. The levels of the starch enzyme brine (SBE) in tuber extracts were measured from 24 transgenic Dianella potato plants transformed with the pBEAll plasmid.
The results showed that transformed transgenic lines of pBEAll produced tubers that have SBE levels that are 10% to 15% of the mvl of SBE found in transformed Dianella plants.
RECAPITULATION
The aforementioned examples refer to the isolation and sequencing of a gene for potato SBE. The examples are further shown that it is possible to prepare antisense constructs of the SBE introns. These constructions can be introduced in the sense of the plants of the SBE to plants, such as potato plants. After the introduction, a reduction in the level of SBE synthesis and / or the level of SBE activity and / or the composition of the starch in the plants can be achieved. While not wishing to be bound by theory, it is believed that the nucleotide sequence of the sense mtron expressed in accordance with the present invention affects the enzymatic activity through co-suppression and / or tsactivation. Reviews of these mechanisms have been published by Fmnegan and McElroy (1994 Biotechnology 12_ pp 883-887) and Matzke and Matzke (1995 TIG 11_ No. 1 pp 1-3). By these mechanisms, it is believed that the sense mte roñes of the present invention reduce the level of enzymatic activity of the plant (in particular the activity of SBE), which in turn for the activity of SBE is believed to influence the ratio from arn? lasa: arn? lopectina and therefore in the branching pattern of Arní Lopectma. Therefore, the present invention provides a method in which it is possible to manipulate the starch composition in the plants, or the tissues or cells thereof, such as potato tubers, by reducing the level of the activity of the plant. SBE using intron sequences of sense. In summary, the present invention therefore relates to the surprising use of antisense vortex sequences in a method to affect enzymatic activity in plants. Other modifications of the present invention will be apparent to those skilled in the art without departing from the scope of the present invention. For example, it is possible to use promoter sequences in anis + sense to affect the enzymatic activity, such as SBE promoter in anis + sense, such as the nucleotide sequence comprising the sequence of nucleotides shown as I.D. SEC. No. 28 to a variant, a depvado or a homologo of the same. The following pages present an amount of axis lists of sequences that have been numbered consecutively from I.D. SEC. No. L - I.D. SEC. No. 29. In a few words, I.D. SEC. No. 1 - I.D. SEC. No. 13 represent sequences of introñes in sense (DNA genónaco) I.D. SEC. No. 14 represents the promoter sequence of SBE (genonac sequence); I.D. SEC. No. 15 - I.D. SEC. No. 27 represents the sequences of tremors in anti-sense; and I.D. SEC. No. 28 represents the sequence complementary to the SBE promoter sequence, ie the promoter sequence of SBE in the orientation an + isentide. The complete sequence of geno- nic nucleotides for SBE including the promoter, the exons and the int roñes is shown as I.D. SEC. No. 29 (see Figures 3 and 8 that highlight- the particular features of the genes). ON SEQUENCES ID. SEC. No. 1 SEQUENCE OF INTRON 1 (1167 bp).
G7- ^ TTR-rACTAATTTCATGt7AATTTCAATTATtTTTAGCCTTrG-ATTTCAtTTTCC- "TATATCT GGA CATC7CCTrAGTTT7TTAT * rTTAT TATAATATCAAATATGGAAGAAAAAtGACAC7TG AG AGCCA7ATG7AAGTA7C ^ TG7sACAAAttr3CAAGGTGGTtsAGTsTAT? -? - AAttCAAAAA7tsAGAGA 7GGAGsGGGGs7GGGGGSAEAGACAA7A777AGAAAGAG7s7TCtAGGAsGttA7GGAGGACACGG 7 AGGGG7AGAAGG77AG77AGG7A7773AG7G77G7C7ssC77A7CCT7TCA7AC7AGTAG_7_CG7GGAA7 7A777GGG7AG77TC77G7777G77A777GA7C777G77A77C7A7T77C7G777Ct7s7AC77CGA77 A7737A77A7A7A7C77G7CG7AG77A77G77CC7CGG7AAGAA7GC7C7AGCA7GC77CC777AG7G7 777A7CA73CC77C777A7A7TCGCG77sC777sAAA7sC777TA r7TAGCCGAGGG7C7A77AGAAA CAA7C7C7C7A7C7CG7AAGG7AG33G7AAAG7CC7CACCACAC7CCAC77G7GGGA77ACA77G7G77 7G77G77G7AAA7CAA77A7G7A7ACA7AA7AAG7GGAT777 7ACAACACAAA7ACA7GG7CAAGGG3 AAAG77C73AACACA7AAAGGG77CA77A7A7G7CCAGGGA7ATGAtAAAAA77G7TrC777G7GAAAG 77A7A7AAGA77tG77A7GGC7777GC7sGAAACA7AA7AAG77A7AA7GC7GAGA7AGC7AC7GAAG7 77G777777C7AGCC7777AAA7G7ACCAA7AA7AGA77CCG7A7CGAACGAG7A7G7777GA77ACC7 GG7CA7GA7G77tC7A7Trtr7ACA7777777GG7G77GAACrGa-A77sAAAA7G7TG7A7CC7A7GA sACGGA7As77GAGAA7s7G7? 7777G7A7GGACC7TGAGAAG-7CAAACGCTAC7CCAAtAA7TrC7A 7GAA77CAAA77CAG77TA7GGC7ACCAG7CAG7CCAGAAAtTAGGA7A7GC7sCATAtAC77G77CAA 7TA7AC7s7AAAA777C77AAG777CAAGA7A7CCA7GTAACC7CGAGAA7TrC77TGACAG
ID. SEC. No. 2 SEQUENCE OF THE INTRON 2 (321 bp)
G7A7G7T7GA7AArr7A7ATGGT73CA7GGA7AG7A7A7AAA7AGT7GsAAAACT7CTGGAC7GG7GC7
CA7GGCA7A777GA7C7G7GCACCG7G7sGAGA7G7CAAACA7GTG7TAC77Cs7TCCGCCAA7TrA7A
A7ACC77AAC77GGGAAAGACAG rC77TACrCCTsTGGsCA77TsT7ATrtsAA77ACAA7CTttA7G
AGCA7sG7G7777CACA77A7CAAC77C777CA7G7GG7A7A7AACAGT777TAGC7CCG77AA7ACC7 T7C77C77777GA7A7AAA rAAC7G7Gs7GCA7TGC77GCBKKK ID. SEC. No. 3 SEQUENCE OF THE INTRON 3 (504 bp). GtAACAGCCAAAAG77stGCT77AGGCAG777GACC77A7ttTGsAAGA7GAA77G7T7A7ACC7AC_ * 7 7GAC77TGC7AGAGAA7777GCA7ACCGGGGAG7AAG7AG7GGC7CCAttTAGG7GGCACC7GGCCA77 7Tt77GA7C7T7TAAAAAGC7G777GATrGGG7C7TCAAAAAAG7AGAaU ^ ^ G7777TGGAGAAG7GAC ACACCCCCGGAG7G7CAG7GsCAAAGCAAAGA7777CAC7AAGGAGA7TCAAAATATAAAAAAAG7A7A GACA7AAAGAAGCTGAGGGGATrC- CA7G7AC7ATACAAGC-A7 jA7ATAG_7_CTrAAAGCAATrrr3 TAGAAA7AAAGAAAG7CTTCC77C7G7TGC77CACAA7TTCCTTCTATTATCA7GAGT7AC7 T7TC7G 7TCGAAA7AGCTTCC77AA7A77AAA7TCATGA7ACTTT7GTTGAGA7TTAGCAG GT7 T TGG7G7A
AAC7GC7C7C7TGGGGGGCAG
ID. SEC. Do not . 4 SEQUENCE OF THE INTRON 4 (146 bp).
GTAGs7CC7CG7C7ACtACAAAA7AstAG7TrCCAtCATCA7AACAsA7TTTCCTATrAAAsCATGATG ttsCAGCA7CA77GGC777C77ACA7GTrC-TAA7TGC7A77AAGG77AtGCTrC7AATTAACTCATCCA CAA7GCAG
ID. SEC. Do not . 5 SEQUENCE OF THE INTRON 5 (218 bp). GTTTTstTATTCA7ACC77GAAGC7GAATTTTGAAC-ACCATC-ATa; CAGGCA7ttCGATTCA7GTtCT7 ACTAG_7_C77GTTA7s7AAGACAt7TrsAAATGCAAAAGTrAAAATAAttGTG7CttTACTAATTTGGAC TTGA7CCCA7A rCTr7CCC7TAACAAAtGAGtCAATTCTATAAGTsCTTGAGAACTTAC A rTCAG CAAT7AAACAG
ID. SEC. No. 6 SEQUENCE OF THE INTRON 6 (198 bp). GTATTTTAAATT7ATr7C7ACAACTA-VATAATTCTC? GAACAATTsp GTCCrG-U-AG7AtAAAAGTACtTAtTTTCGCC-ATGsGCCtTCAG- ^ TATTGGTAGCCsCTGAATATCAt
GATAAGTrATTTATCCAG7GACATTTtTATGTTCACTCCrAtTATGTCTGCTsGATACAG ID. SEC. No. 7 SEQUENCE OF THE INTRON 7 (208 bp).
GTtTGTC7G7TrctA77GCATT77AAGGTrCATATAGG7TAGCCACsGAAAAtCTCACT rtTGTGAGG 7- ^ CCAGGG7TC7GA7GGATTA77 AAT C CsTTrATCA rrG? Tli.A? CT TCATGCATTGTG7 pc? Tr? c 7Atccc7: rAttr5- ^ GGtAATi? ttCT
ID. SEC. No. 8 SEQUENCE OF THE INTRON 8 (293 bp).
GTATGTCT7ACA7C r7AGATA777TGTGATAArrACAATTAGTT7GGCTrAC-TTGAACAAGAT CAT7 CCTCAAAATGACCTG- C7GTTGAACATC-UlAGGGGTTGAAAC-ATAG-U-GAAAA ülC-ATGA7GAATG7
TTCCATTGTCTAGGGA77T - TATTATGTTsCTGAGAACAAATGTCATCTTAAAAAAAACATTGTTTAC7 TtTTTGTAGTATAGAAGATTACTG7ATAGAGtTTGCAAGtsTGtCTUTrp, GGAGTAATTs7GAAATsT TTGA7GAACTTGTACAG
ID. SEC. No. 9 INTRON 9 SEQUENCE (376 bp). GTTC-UVGTATTTTGAA7CGCAGCTTstTAAATAATCTAGtAATT- ^ cttGGTTCTsssGA GA AGCTCAtttcAtcptsttCTACttAtrrTC tTrCGAGAtCC_AAGtATTAGAp - ATTtACACTtATtACCGCCTC AGCAGCrtAAGTTGA7TCrTT - AAGCTATAGTTTCAGGCTACC-AATCC ^ ATACT7ACCTTTTCTT7AC-AA7GAAG7sA7AC7AATTGAAA7GsTC7AAA7C7sA7A7C7A7ATTTC7C
CGTCTTTCCTCCCCC7 A7GA7GAAA7GCAG ID. SEC. No. 10 INTRON 10 SEQUENCE (172 bp).
GtAAAA7CA7C7AAAG77sAAAG7G77GGG777A7GAAG7GC777AA7TC7A7CCAAGGACAAG7AGAA ACC77Tr7ACCtTCCA777C77GA7GA7GGA777CATAT7A77TAATCCAA7AGC7GG7CAAA77CGG7 \ 7AGC7G7AC7GAT7AGT7ACT7CACT77GCAG. 11 SEQUENCE OF THE INTRON 11 (145 bp).
GTA7A7A7s77TTAC77A7CCA7GAAA77A77GC7C7GC7TGTI T7AA7s7AC7GAAC_AAG7777A7G GAGAAG7AAC7GAAA ^ AA7CA7777CACA77G7C7AA777AAC7CTr: "r77C7GA7CC7C3CA7GACG AAAACAG
ID. SEC. No. 12 SEQUENCE OF THE INTRON 12 (242 bp).
GTAAGGA7TrGC77GAA7AACT'r: ':, GA7AA7AAGATAACAGA7G7AGGG7ACAG77C7C7CACCAAAAA GAAC7G7AA7TG7C7CA7CCA7C777AGTrs7AtAAGA7AtCCGACTGtC7GAG77CGGAAG7G77TGA GCC7CC7GCCC7CCCCC7GCG77G777AGC7AA77CAAAAAGGAGAAAAC7s777A77GA7GA7CT 7G 7CTTCA7GC7GACA7ACAA7C7G77C7CA7GACAG
ID. SEC. No. 13 SEQUENCE OF THE INTRON 13 (797 bp).
? GtACAGTrC77GCCs7G7GACC7CCC7TT77A77GTss777TG77CATAG_77_A777GAA7GCGA7AGAA s77AAC7A77GA7TACCGCCACAA7CsCCAG77AAG7CC7C7GAAC7AC7AA777GAAAGs7AGGAA7A 5 GCCG7AA7AAGG7C AC77T7GGCA7rr7ACrGtTACAAAACAAAAGGATsCCAAAAAAA77C77C7C7 ATCC7C7T777CCrrAAACCAG7GCAtG7AGC77sCACCTsCATAAAC7TAGG7AAA7GA7CAAAAA7G AAG77GA7GGGAAC7TAAAACCGCCCtGAAG7AAAGC-tAGGAATAGTCATATAA7G7CC CCTT7sGTG 7C7GCGCTAACA7C- ^ C-AACAACA7ACCtCG7G7AGtCCCACAAAGTGGTTtCAGGGGGAGGG7AGAG7 GTATGC -? \ AAC-TAG-ACCC77GGC7C- TrACTCCTA7C7CAGAGs7AsAGAGGA7TTTTrC- VGAAA AAGTCCAAAAAGAAGtAA - AGAAG7GAAAGCAACATGTGTAGCTAAAGCGACCCAAC77G7TtGGGAC7 GAAG7AG7tsrrsTTs7tsAAACAs7GCA7G7AGATGAACACAtGTCAGAAAA7GGACAACACAGTrA7 7TrG7GCAAGtCA- -AAAA7GTAC7AC7AT i T'r GTGCAGCtTTATGTA7AsAAAAG7TAAA7AAC7 l (] AATGAATrTTGCtAGCASAAAAA7AGC7TGGAGAGAAArrrttTATATrGAAC7AAGC7AAC7A7A77C ATCTT7CTT7TTGCTTCTTCTTC7CCTTGTT7G7GAAG
fifteen
twenty
ID. ? EC. No. 14 DNA SEQUENCE OF THE PROMOTING REGION OF THE SBE GENE
ATCATGGCCAA77AC7GG77C ?? A7sCAT7ACtTCCTT7CAGATTCTTTCGAG77C7CA7 60
GACCGG7CC7AC7? CAG? CGA7AC7AACCCs7GGAACT3TTGCA7C7GCTTCr7AGAAC7 120
CTA7sGC7A7T rCG77AGC773GCG7CGGTr7sAACA7AGTT7TTG77TTCAAAC7CTT 180
CA7TTACAG7CAAAA7G77G7A7GG7t77TG77TTCC CAATGA7G7TrACAG7G77stG 240
7TGTCA7C7G7AC7TT7GCC7A77AC7TG? N? TTGAG7TACAtGTTAAAAAAGtG777AT7 300 psCCA7AT-TtGT7C7C77A77A77A7TA7CATACATACATTATTACAAGGAAAAGACA 360
AG7ACACAGA7C77AACG777A7s77CAA7CAAC7TrTGGAGGCATrGACAGG7ACCACA 420
AArr7TGAG7TrA7GA77AAG77CAA7C77AsAATATGAATTTAACA7CTATrA7AGA7G 4B0
CATAAAAA7AGC7AA7GA7AGAACATrsACATrTGsCAGAGCTTAGGG7ATsG7A7A7CC 5 0
AACs77AATr7AG7AA77777s77ACG7ACG7A7A7GAAATA7TGAArrAA7CACA7GAA 600
CGG7GsA7A77A7ArrA7GAG77GGCATCAGCAAAATCATrssTG7AG7TsAC737AGTr 660
GCAGA7t7AA7AA7AAAA7GG7AArrAACGG7CGA7ATTAAAAtAAC7CTCA7T7CAAG7 720
GGsA77AGAAC7As77A77AAAAAAA7G7A7AC7T7AAG7GAtTTGATGGCATA7AA7t7 780
AAAG7T777CA7TrCA7GC7AAAATtG77AATtATtGTAATsTAGACtGCGAC73GAA77 8 0 ATtA7AG7s7AAATr7A7GCA77CAG7s7AAAAttAAAGtATTGAACT7GTC7s777tAG 900 AAAA7ACTr7A7AC777AA7A7AGGATttrG7CAtGCGAATTTAAArrAATCGA7A7TGA 960
ACACGGAA7ACCAAAA77-? AAAGGATACACA7GGCCTTCAtATGAACCG7GAACC777s 1020
ATAACG7GGAAG77CAAAGAAGs7AAAG77TAAGAATAAACTsACAAATTAATr7C7TT7 X0B0
ATTTGGCCCAC7ACGAAA7773C777AC7TTCTAACATGTCAAGTTGTGCCCTC77AG7T 1140 sAA7GA7A77CA7TTrTCA7CCCA7AAGTTCAATTTsATTsTCAtACCACCCA7GA7G7T 1200
CTG-UU «ATsCTrGGCCA77CAC-AAAG77TA7CTTAG_7_TCCtATGAAC7TTATAAGAAGC 1260
TT7AATTrGACA7GTrA777A7A77AGATGA7ATAAtCCATsACCCAATAGACAAG7G7A 1320
TrAA7ATTG7AAC7tTG7AA77GAG7G7G7C7ACA7CTTAtTCAA7CA7TrAAGG7CA77 1380
AAAA7AAATrA777'¿ "r7GACA77C7AAAACT7TAAGCAGAATAAATAG_7_TTA7CAA7tA7 1440
7AAAAACAAAAAACGAC77A777A7AAA7CAACAAACAATtt7AGA77GCTCCAACA7A7 1500
7ttTCC -? - ATrAAA7GCAGAAAA7GCA7AATr77ATACTTGATCTrrATAGCT7A77Tr7 1560 tTTAGCCT- ^ CCAACs- ^ 7A777G7AAACrCAC- ^ CT GAttAAAAGGGAtt7ACAACAA 1620
GAtATAtA7AAG7AGTOACAAA7C7TGATr7TAAATATrTTAA7TTsGAGGTCAAAA777 1680
TACCATAATCA7TtG7A777ATAATTAAATTTT -? - ATATCttATTTATACATATC7AG7A 1740
? AAC7t AAATAtACG7A7A7ACAAAAtATAAAATTATTGGCstTCATATrAGGtCAA7A 1800 AAtCCtTAACTATATC7GCCTtACCACTAGGAGAAAGTAAAAAACTCTTTACCAAAAATA CATGTAtTATGTAtA-AAAAAGtCGATTAGAtTACCT 1860 - ?? TAGAAATTGTATAACGAGTA AGTAAGTAGAAAtATAAAAAAACTAC-AATACTAAAAAAAATATGtTTTACTtCAA7TtCG 1920 1980 AAA T- TGGGGTCTGAGTGAAATATTC-AG- ^ V GGGGAK -? ACTAACAAA GGGTCA7AA7 GTT77TTTA7AAAAAGCCACTAAAATGAGGAAA7CAAGAATCAGAACATACAAGAAGGCA 2040 2100 GCAGCTGAAGOVAAGTACCAT- TTTAATt --- ^ TGGAAATTA 2160???
ACCCATTCG ID. SEC. No. 15 INTRON 1 ANTICIPATION SEQUENCE (1167 bp). CTG7CAAAGAAA77: CGAGG77ACATGGATATCrtsAGAA-Tr-? IsAAATrtrACAG7A7AATtGAACGCA7A7CC7AA777CtsGACtGACTGG7AGCCATAAAC7GAA777GAATrCATAGAAA
TTATTGsAG7AGCG777GAGC77CT - AAGGTCCAtACAAAGAACACA77C7CAAC7A7CCGtCTCATAG
GATACAACATr7TC-? 77GCAG77CAACACCAAAAAAATG7AAAAAATA-y ^ UCATCATGACC-AGGT ^
7C- AACA7AC7CG77CGA7ACGGAA7CTATTATTGGTACAtTTAAAAGGCrAGAAAAAACAAACTTCA
GTAGC7A7C-7CAGCA77A7AACTrATtATGTTTCC_AGC-AAAAGCCATAAC ^ C-U-AGAAACAA77777A7CATA7CCCTGGACATATAATGAACCCTTrATGTG77CAGAACTrTGCCCTT
GACCAts7A7T7G7G77stAAAAAATCCACtTATTATG7ATACATAA7TGA7ttACAACAACAAACACA
ATGTAA7CCCACAAG7sGAG7G7GG7GAGGAC7TTACCCCtACCTTACGAGATAGAGAGAt7sTTrC7A
ATAGACC rCGGC7-y-AG7AAAAGCATtTCAAAGCAACGCGAA7A7AAAGAAGGCA7GA7AAAACAC7A
AAGGAAGCA7GC7AGAGCA77C77ACCGAGGAACAAtAACTACGACAAGA7A7A7AA7ACAATAATCGA
AG7ACAAGAAACAGAAAA7AGAA7AAO VAGA7C-AAATAACAAAA-lAAGAAAC7ACCCAAA7AATrCCA
CGAC7AC7AGTA7GAAAGGA7AAGCCAGA -ACACTCAAATACCTAAC7AACC77C7ACCCCTCATCCG
7GTCC7CCA7AACCTCC7AGAAC-AC7C7TTCTAAAtArrsTCT? * TVCCCCCACCCCCCC7CCATCTC7C
AAttrT7GAATTrrA7ACACTCAAC-ACCttGCAAATTTGTCACATGA7AC TACATA7GGCTCTACAA sTGTCA7T77TC77CCA7ATTTGA7ATTATAAAAAATAAAATAAAAAACTAAGGAGATGATCCAGATAT
ATTGGAAAA7GA? A7GCAAAGGCGAAAAATAATTGAAATTAACA7GAAATTAG_7_AAAA? TTAC
ID. SEC. No. 16 INTRON 2 ANTICIPATE SEQUENCE (321 bp).
MMMVGCAAGCAATGCACCAC-AGTrAGTTTAtATCAAAAAGAAGAAAGGTAttAACGsAGCtAAAA rrA7ATACCACA7sAAAGAAG77GA7AATG7G-W ^ \ AACACCA7s rCA7AAAGA7Ts7AA77CAAA7AAC AAATGCCCACAGGAG7AAAGAGCTGTCTITCCC-UGTTAAGsTATTATAAATTssCGGAACsAAGTAAC ACAtGTTrGACA7C7CCACACGG7GCAa ^ AtCAAATATGCCATGAGCACCAG7CCMAAGTTrTCC-AA
CTAttTATATACTA7CCA7GCAACCATATAAAtTATCAAACATAC
ID. SEC. No. 17 INTRON 3 ANTICIPATION SEQUENCE (504 bp).
CTGCAAAAAAAGAGAGCAG7TTACACAAGAAAAAACTGC7AAATCTCAAC-AA? AGTATC-A7GA? RrT-7ATTAAGGAAGCTATT7CGAACAGAAAGAGTAACTCATGATAATAGAAGGAAATTGTGAAGCAACAGAA GGAAGA l 'CTllA T' ACA-U-AT ü "rAA --- A rA7A T - A * G? TGTATAG_7_ACA'rü GAA
7CccctcAGcttctttAtstctATAc rr rp'ATAr?, R ü- ^ t rccttAGTGAAAAtcttto, n, G
CCACTGACACTCCGGGGGTGTGTCACTTCTCCAAAAACCTTGrTCTACTTTTTTGAAGACCCAATCAAAC AGCT7TTTAAAAGA7CAAAAAAA7GGCCAGGTGCCACC AAATG - AGCCACTACTTAC7CCCCGGTATG CAAAATtCTCTAGCAAAGTCA- G7AGGTATAAACAATT tCTrCCAAAATAAGGTCAAACT ^ AGCACAAC1T7TGGC GTTAC?
ID. SEC. No. IB SEQUENCE IN ANTICIPATION OF THE INTRON 4 (146 bp).
CTGCATTG7GGA7GAG7TAA7TAG-? GCA7AACCTTAA7AGCAATTAGAACA7G7AAGAAAGCCAATGA
7GC7GCAACA7CA7GC77TAA7AGGAAAA7C7G77ATGATGA7GGAAACTAC7AtT7TG7AGTAGACGA GGACCTAC
ID. SEC. No. 19 INTRON 5 ANTI-SEQUENCE SEQUENCE (218 bp). CTG77TAA77GC7sAAG7AG7AAG7TCTCAAGCAC7TATAGAATTsACTCATTTTs7TAAGGGAAAGAG 7A7GGGA7CAAGTCCAAATTAG_7_AAAGAC? CAAtTATTTr- CttTTGCATTrCAAAA7s7C7TACATA ACAAGACrAGTAAGAACA7GAATCGAAA7GCCTG7GA7GATGGtGTrCAAAArrCAGC77aVAGGTA7G AATAACAAAAC
ID. SEC. No. 20 INTRON 6 ANTICIPATION SEQUENCE (198 bp).
C7G7A7CCAGCAGACA7AATAGGAG7GAACA7AAAAATG7CACtssA7AAA7AAC77A7CA7GATAt7C AGCsGCTAC3AATATrC7s- GGCCCATsGCGAAAATAAG7ACTTTrAtACTTTCAGGACGTATAt * - "• 7GGA77C7ATCTAACAA7tGTtC7GAGAA7TA7TTAsTTGTAGAAATAAA7tTAAAA7AC
ID. SEC. No. 21 INTRON 7 ANTICIPATION SEQUENCE (208 bp).
CTG7GG77AGAAGC7-U-AAGtsAA7AGAtGAGAAAAATrACCtCCAAATAAGAGGGA7A77sAAAAAGA AACACAA7GCATGAAAAGAATAAACAAAtGA7AAACsAGAAAATTO-? T ^ TCCATC-AGAACCCtGG77
ACC7CACAAAGAG7GAGA7TttCCG7GGC7AACC7ATATGAACC TAAAA7Ga? TAGAAACAGACAAAC
ID. SEC. No. 22 INTRON 8 ANTICIPATION SEQUENCE (293 bp).
CTGtACAAGTrCA7CAAACATrtCACAATtACrCCAAAA? GAC-ACACTTsCA- TC7ATAC7ACAAAAAAGTAAACAATÜlTlTl ^ C? N'AAGATGACATrTG7TC-TCAGCAACAtAATAGAA
ATCCC7AGACAAtGGAAAC? TTCA7CAtGTTGT7TrCCTCTATGTTTCA TrCAGGTCA rr ^^ ^ tGAA7TtGTTC- AGG- GT-U 3CCAAACTAATTGTAATtATCACAA ^
AAAGA7G7AAGACA7AC
ID. SEC. DO NOT . 23 ANTICIPAL SEQUENCE OF THE INTRON 9 (376 bp). CTGCATT7CATCA7GAGGGGGAGGAAAGACGGAGAAATA7AGA7ATCA - AT77? GACCATT7CAATTAG_7_A7CACT7CATTG7-U-AGAAAAGG7AAG7A7CCAACAAA7A7AGC - AGGCT C7A7AGCT7CAAAGAA7C-? CT7AAGCTGC7CA7C - U-GGCCT7AGTsG7AGA TT AAATGAATC A7ACTtGGA7C7CsAAACAAAAATCA3AAATTCGG77sGAAAA7AAs7A £ -J-A (- AA
GA7GAAA7GAGC7A7CA7CCCCAG-? CC- ^ G7AGA r7CCAAG7AAGCAA7C7AAAAA77AC7AGA77A
777AACAAGC7GC3A77CAAAA7AC77GAAC
ID. SEC. Do not . 24 INTRON 10 ANTI-SEQUENCE SEQUENCE (172 bp).
C7sCAAAG7GAAG7AAC7AA7CAG7ACAGC7A77ACCGAA777GACCAGC7A77GGA77AAA7AATA7G AAA7CCA7CA7CAAGAAA7GGAAGG7AAAAAGs777C7AC77G7CC77GGA7AGAA77AAAGCAC77CA 7AAACCCAACAC777CAAC777AGA7GA7777AC
ID. SEC. No. 25 INTRON 11 ANTI-SEQUENCE SEQUENCE (145 bp).
C7s7777CG7CA7sCGAGGA7CAGAAAAAAGAG77AAA77AGACAA7G7GAAAA7GA777G777CAG77 AC77C7CCA7AAAAC77377CAG7ACA77AAAAACAAGCAGAGCAA7AA777CA7GGA7AAG7AAAACA 7A7A7AC
ID. SEC. No. 26 INTRON 12 ANTICIPATION SEQUENCE (242 bp). CrG7CA7GAGAACAGA77G7ATs7CAGCATGAAGAC-AAAGA7CA7C- 7AAACAG7777C7CC7tTrrG AAT7AGC7AAAC- ^ CGCAGGGGGAGGGCAGGAGGCTCAAACAC-T7CCGAACTCAGACAGTCGGATATC7 7A7AC - U.CrAAAGA7GGA7GAGACAAtTACAGri rn l, ÜG7GAGAGAA rs7ACCC7ACA7 rG77A
7CT7AT7A7CAAAAGT7ATtCAAGCAAATCCTTAC ID. SEC. No. 27 INTRON 13 ANTICIPATION SEQUENCE (797 bp).
CT7CACAAACAAGGAGAAGAAGAAGCAAAAAGAAAGA7GAA7A7AG77AGC77AG77CAA7A7AAAAAA 7TrC7C7CC-? GC7A77777CTGC7AsCAAAATrCATrAG77A777AAC7777C7A7ACATAAAGC7GC ACAAAGAAA7AG7AG7ACA7TT7777GAC77GCACAAAA7AAC7G7G77G7CCA7777C7GACATG7G7 7CA7C7ACA7sCAC7G777CAACAACAACAAC7ACT7CAG7CCCAAACAAGTrsGG7CGCTr7AGC AC ACA7G77sC777CAC77C7GT7AC'i "r TTTGGA, rp, T TC? 7GAGCCAAGGGTC7AT7GAAAAAA 7CC7C7C7ACC7C7GAGA7AGGAG7AAsTT77GCA7ACAC7C7ACCC7CCCCCTGAAACCAC rTs7GG
AGC7TTAC77CAGGGCGGTTTrAAG7TCCCATCAA ITCATrtr7GA7CA77TACC7AAG7TrATGCAG GTGO? GC7ACA7GCACrGG777AGGGAAAAAGAGGATAG - «- AAGAA777TT7TGGCA7CC7TttGT77
7G7AACAG7AAGA7GCCAAAAG7AGACCTGA77ACGGC7A77CC7ACC777CAAA77AG7AG77CAGAG GAC77AAC7GGCsA77s7GGCGG7AA7CAA7AG77AAC77C7A7CGCA77CAAA7AAC7A7sAACAAAA CCACAATAAAAAGGGAGG7CACACGGCAAGAAC7GTAC
ID. SEC. No. 28 SEQUENCE OF ANCIENT DNA IN THE REGION PROMOTING THE GENE
SBE. CGAA7GGG7777GA7AAAACTrrGAAA77AA7TrCCA77GA77AAA77ATGG7AC777GC 60
T7CAGC7GC7GCCT7C7TG7A7GT7CTGAT7CTTGATTrCC7CATTT7AG7GGCTTTT7A 123
7AAAAAAACA77A7GACCCT7T 3T7AG7CC7CCCCTGGCTGAA7A777CAC7CAGACCC 180
CArrAsTT7CGAAAT7GAAG7AAAACA7ATTTTTTrrMTATTG7AGTTTTrT7ATATT7 240
CTAC-TrAC77AC7CG77A7ACAA777CTATrrAGGTAA7CTAA7CGACTTrr7G7A7ACA 300
7AA7ACA7G7AT7777GG7AAAGAG'1"; TTTTACT77C7CC7AG7GG7AAGGCAGA7A7AG 360
77AAGGA777A7tGACC7AATA7GAACGCCAA7AA7TT7A7A7777G7A7A7ACG7ATA7 420
77AAAAG777AC7AGA7A7G7A7AAA7AAGA7A777AAAA7TGAA77ATAAA7ACAAA7G 4B0
A77A7Gs7AAAA7777GACC7CCAAA77AAAA7ATrrAAAA7CAAGA7TrG7CAC7AC77 540
ATA7A7A7C77G77s7AAA7CCC7777AA7CAAG77G7GAGTr7ACAAA7A77CG77GG7 600
7AGGC7AAAAAAAA7AAGC7ATAAAGA7CAAG7A7AAAA77A7GCA7777C7GCA777AA 660
TrrsGAAAAA7A7G77GsAGCAA7C7AAAA77sTr7GTrGAT77A7AAA7AAG7CG7777 720
T7GTTTT7AA7AAT7GA7AAAC7ATTTATTC7GCTGAAAGTGTGAGAA7G7CAAAAAA7A 780
Arr7A7777AA7GACC77AAA7sA77GAA7AAGA7G7AGACACAC7CAA7tACAAAG77A 840
CAA7ATrAA7ACAC77G7C7ATrsGGTCA7GGA77A7ATCA7C7AA7ATAAA7AACA7G7 900
CAAA77AAAGC77C77A7AAAG77CA7AGGAAC7AAGA7AAAC7TGG7GAA7GGCCAAGC 960
ATTrn'CAGAACA7CA7GGG7GG7A7GACAA7CAAA77GAAC77A7GGGA7GAAAAA7GA 1--20
A7A7CA77CAAC7AAGAGGGCACAACttGACA7strAGAAAG7AAAGCAAA7ttAG7AG7 1080
GGsCCAAA7AAAAGAAA77AA777G7CAG7T7A C'7AAAC777ACCT7CT7TGAAC77 1140
CCACG77A7CAAAGG77CACGG77CA7ATGAAGGCCATsTGTA7CCTTTTTAA rTGG7 1200
AttCCG7G77C- ^ 7A7CGATTAA77TAAATrCGCA7GACAAAA7CC7AtAtrAAAGtA7A 1260
AAG7A7777C7A-W? CAGAC-AAG77C -? - 7AC7TrAAT7TrAC-AC7s-? 7sC_A7AAATrrA 1320
????? CAC7A7AA7AA77CCAG7CGCAG7CTACA7TACAA7AATGAACAA77TGAGCA7GAAA7G AAAAAC777AAA77A7A7GCCA7CAAA7CAC77AAAG7A7ACAT7777 1380, p, AA7AAC7AG7 7C7AA7CCCAC77GAAA7GAGAG77A77TGAA7A7CGACCG77AA77ACCA777TA77A7 1440 1560 1500 7AAA7C7GCAAC7ACAG7CAAC7ACACCAA7GAT777GC7GA7GCCAAC7CA7AA7A7AA 7A7CCACCG77CA7G7GA7TAA77CAATATrrCATATACs7ACG7AACAAAAA7tAC7AA AtTAACG77GGA7A7ACCA7ACCC7AAsC7C7sCCAAA7GTCAA7GTrCTA7CA7TAGC7 1620 1680 1740 CC AAA777G7GG7ACC7G7CAA7GCC7CCAAAAG77GA77GAACA7AAACG77AAGA7 ATT7'i A7sCA7C7A7AA7AsA7G7tAAA7TCATAtTCTAAGATrGAACt7AATCA7AAA 1800 CTG7G7ACTrGTC r7TCC7TG7AA7A TGTATGTA7G TAATAA7AATAAGAGA A7A7GGCAAAA7AAACA-Trr777AACA7G7AAC7CAAAACAAG7AA7AGGCAAAAG7AC CAAA 1860 1920 1960 AGA7GAC ACACAACACTGTAAACATCATTGAGGAAAAC-AAAAACCATACAACATTrrGA CTG7AAA7GAAGAG777G - - AA ( --AAAAA - 7A7G77C -? - ACCsACsCCAAGCrr ^ 2040 7AGC-A7AGAstrC7AAG? AsCAGAtsC-UCAGTTCCACsGGTrAGtA7CGTCTGtAGTA 2100 GGACCGG7CA7G? GA? C7CGAAAGAATCTGAAAGGAAG7AATGCA777GAACCAGTAAT7 2-L60 GGCCA7GA7 SEC. ID No. 29 GEN SBE GENOMICA77AC7GGT7 CAAA7GCA77 ACTTCCTT7C AGATTCTTTC GAG7TCTCA7 60
GACCGG7CC7 AC7ACAGACG A7AC7AACCC G7GGAAC7G7 7GCA7CTGC7 7CTTAGAAC7 120
CTATGGC7A7 777CG77AGC 77GGCG7CGG TTTGAACATA GT T'-. ITT 7CAAACTCTT 180 CATTTACAG7 CAAAA7G77G TATGGTTm 'GTTTTCCTCA ATGA7G7T7A CAGTGTTGTG 240 ttstcA7C7s TACTTTGGCC 7ATGACTTG7 TTTGAGTTAC ATGT7AAAAA AGTGTTTATG 300
7TGCCA7A77 77G77C7C77 AT7AtTA7TA 7CATACATAC ATTA77ACAA GGAAAAGACA 360
AGTACACAGA 7C77AACG7T 7A7G7TCAA7 CAACTTTTGG AGGCATTGAC AGGtACCACA 420
AATTTTGAG7 77A7GA77AA GT7CAATCT7 AGAA7AtGAA 7T7AACA7C7 ATtATAGA7G 480
CATAAAAA7A GC7AA7GA7A GAACATTGAC A7T7GGCAGA GC7TAGGG7A 7GGTATA7CC 540
AACG77AA77 7AG7AA7777 7G77ACG7AC G7A7A7GAAA 7A7TGAA77A A7CACA7GAA 600
CGGTGGA7A7 7A7A77A7GA G77GGCATCA GCAAAATCA7 7GG7G7AGTT GAC7G7AG77 660 GCAGA777AA 7AA7AAAA7G G7AA7TAACG G7CGA7ATTA AAA7AACTC7 CAT7TCAAG7 720
GGGA77AGAA C7AG77A77A AAAAAA7G7A 7ACTT7AAG7 GA777GATGG CATATAA777 780
AAAGTTTT7C ATT7CA7GC7 AAAA77G7TA A7TATTGTAA 7G7AGAC7GC GACTGGAA77 840
ATTATAG_7_G7 AAA777A7GC AT7CAGTGTA AAATTAAAG7 AtTGAACTTG 7C7GTT7TAG_900_AAAA7AC777 ATACT77AA7 A7AGGA7T77 GTCATGCGAA 7TTAAATTAA TCGATATTGA 960
ACACGGAA7A CCAAAA77AA AAAGGA7ACA CATGGCCT7C ATATGAACCG TGAACCtTTG 1020
ATAACG7GGA AGT7CAAAGA AGG7AAAGT7 7AAGAATAAA CTGACAAATT AATGGCTTTG 1080
ATTTGGCCCA CTACTAAATT 7GCTTTACT7 7CTAACATGT CAAGTTGTGC CCTCTTAGTT 1140 GAA7GA7A77 CATTTTTCAT CCCATAAGTT CAATTTGATT GTCATACCAC CCATGATGTT 1200
CTGAAAAATG CT7GGCCA77 CACAAAGTT7 ATCTTACTITC CTATGAACTT TATAAGAAGC 1260
TTTAATTTGA CA7GT7ATT7 A7ATTAGA7G ATATAATCCA TGACCCAATA GACAAGTGTA 1320
TTAATATTGT AAC777G7AA 77GAGTGTG7 CTACATCTTA TTCAATCATT TAAGGTCAT7 1380
AAAATAAAT7 ATTT7T7GAC A7TCTAAAAC TTTAAGCAGA ATAAATAGTT TATCAATTAT 1440
TAAAAACAAA AAACGACTTA 77TATAAATC AACAAACAAT tTTAGATTGC TCCAACATAT 1S00
TTTTCCAAAT TAAATGCAGA AAATGCATAA TTTTA7AC7t GATCTTTATA GCTTATTTT: '1560
TTTAGCCTAA CCAACGAATA TTTGTAAACT CACAACTTGA TTAAAAGGGA TTTACAACAA 1620 GATATATATA AGTAGTGACA AATCTTGATT TTAAATATTT TAATTTGGAG GTCAAAATTT 1680
TACCATAATC ATTTGTATTT ATAATTAAAT TTTAAATATC TTATTTATAC ATATCTAGTA 1740
AACTTTTAAA TATACGTATA TACAAAATAT AAAATTATTG GCGTTCATAT TAGGTCAATA B 00
AATCCTTAAC TATATCTGCC TTACCACTAG GAGAAAGTAA AAAACTCTTT ACCAAAAATA 1860
CATGTAT7A7 GTATACAAAA AGTCGATTAG ATTACCTAAA TAGAAA77G7 ATAACGAGTA 1920 AGTAAGTAGA AATATAAAAA AACTACAATA CTAAAAAAAA TATGTTTTAC TTCAATTTCG 1980
AAACTAATGG GGTCTGAGTG AAATATTCAG AAAGGGGAGG ACTAACAAAA GGGTCATAAT 2040
GTTTTTTTTAT AAAAAGCCAC 7AAAATGAGG AAATCAAGAA TCAGAACATA CAAGAAGGCA 2100
GCAGCTGAAG CAAAGTACCA 7AATTTAATC AATGGAAATT AATTTCAAAG TTTTATCAAA 2160
ACCCATTCGA GGATCTTTTC CATCTTTCTC ACCTAAAGTT TCTTCAGGGG TAATTTTTAC 2220
TAATTTCATG TTAATTTCAA TTATTTTTAG CCTTTGCATT TCATTTTCCA ATATATCTGG 2280
ATCATCTCCT TAGTTTTTTA T7TTATTTTT TATAATATCA AATATGGAAG AAAAATGACA 2340
CTTGTAGAGC CATATGTAAG 7ATCATGTGA C AATTTGCA AGGTGG77GA GTGTATAAAA 2400"r" AAAAATT GAGAGATGGA GGGGGGGTGG GGGBARAGAC AATATTTAGA AAGAGTG77C 2460 GGTTA TGGAGGACAC GGATGAGGGG TAGAAGGTTA GTTAGGTATT TGAGTGTTGT 2520
CTGGCT7A7C CTT7CA7AC7 AG7AG7CGTG GAATTAT7TG GGTAG_7_TTC7 TGTTTTG7TA 2580
TTTGATC7TT GTTA77CTAT 777C7GTTTC TTGTACTTCG ATTATTGTA7 TATATATCTT 2640
G7CGTAG_77_A 7TGT7CC7CG G7AAGAA7GC 7CTAGCATGC TTCC777AG7 GTTTTATCAT 2700
GCCTTC777A 7A77C3CG77 GC777GAAA7 GC7777ACTT TAGCCGAGGG 7CTATTAGAA 2760
ACAATC7C7C 7A7C7CG7AA GG7AGGGG7A AAG7CC7CAC CACAC7CCAC 77GTGGGATT 2820
ACA77G7G7T 7G77G7TGTA AATCAAT7A7 GTATACA7AA 7AAGTGGA7T 7TTTACAACA 2880
CAAATACA7G G7CAAGGGCA AAG77C7GAA CACA7AAAGG GTTCATTATA TGTCCAGGGA 2940
TATGATAAAA AT7GT77C777G7GAAAG77 ATATAAGATT TGTTATGGC7 TTTGCTGGAA 3000
ACATAATAAG T7A7AA7GC7 GAGATAGCTA CTGAAGTTTs TTTTTTCTCG CCTTTTAAAT 3060
GTACCAATAA TAGA77CCG7 A7CGAACGAG TATGTT7TGA TTACCTGGTC ATGATG77TC 3120
TATTTTTTAC ATGGTTTTGG 73T7GAAC7G CAATtGAAAA TGTTGTATCC TATGAGACGG 3180
ATAGTTGAGA ATGTG77C777G7ATGGACC TTGAGAAGCT CAAACGCTAC TCCAATAATT 3240
TCTATGAATT CAAAT7CAG777A7GGC7AC CAGTCAGTCC AGAAATTAGs ATATGCTGCA 3300
TATACTTGTT CAATTATACT GTAAAATTTC TTAAGTTCTC AAGATATCCA TGTAACCTCG 3360
AGAAT7TCTT TGACAGGCTT C7AGAAA7AA GATATG7TTT CCTTCTCAAC ATAGTACTGG 3420
ACTGAAG7T7 GGATC7CAGG AACGGTC77G GGA7A77TCT TCCACCCCAA AATCAAGAGT 3480
TAGAAAAGAT GAAAGGG7A7 G777GA7AA77TATATGG7T GCA7GGATAG TATATAAATA 3540
GTTGGAAAAC TTCTGGACTG G7GCTCATGG CATATTTGAT CTGTGCACCG TGTGGAGATG 3600
TCAAACATGT GTTACTTCG77CCGCCAATT TATAATACCT TAACTTGGGA AAGACAGCTC 3660
TTTACTCCTG TGGGCATTTG 7TATTTGAAT TACAATCTTT ATGAGCATGG TGTGTTCACA 3720
TTATCAACTT CTTTCATGTG GTATATAACA GTTTTTATSCT CCGTTAATAC CTTTCTTCTT 3780
TTTGATATAA ACTAACTGTG GTGCATTGCT TGCBíaOATG AAGCACAGTT CAGCTATTTC 3840
CsCTGTTtTG ACCGATGACG ACAATGCGAC AATGGCACCC CTAGAGGAAG ATGTCAAGAC 3900
TGAAAATATT GGCCTCCTAA ATTTGGATCC AACTTTGGAA CCTTATCTAG ATCACTTCAG 3960
ACACAGAATG AAGAGATATG TGGATCAGAA AATGCTCATT GAAAAATATG AGGGACCCCT 4020
TOAGGAATTT GCTCAAGGTA ACAGCCAAAA GTTGTGCTTT AGGCAGTTTG ACCTTATTTT 4080
GGAAGATGAA TTGTTTATAC CTACTTTGAC TTTGCTAGAG AATTTTGCAT ACCGGGGAGT 4140
AAGTAGTGGC TCCATTTAGG 7GGCACCTGG CCATTTTTTT GATCTTTTAA AAAGCTGTTT 4200
GATTGGGTCT 7CAAAAAAG7 AGACAAGGTT TTTGGAGAAG TGACACACCC CCGGAGTGTC 4260
AGTGGCAAAG CAAAGATTTT CACTAAGGAG ATTCAAAATA TAAAAAAAGT ATAGACATAA 4320
AGAAGCTGAG GGGATTCAAC ATGTACTATA CAAGCATCAA ATATAGTCTT AAAGCAATTT 4380 TGTAGAAATA AAGAAAGTCT 7CCTTCTGTT GCTTCACAAT TTCCTTCTAT TATCATGAGT 4440
TACTCTTTCT GTTCGAAATA GCTTCCTTAA TATTAAATTC ATGATACTTT TGTTGAGA77 4500
TAGCAGTTTT TTCTTGTGTA AACTGCTCTC TTTTTTTGCA GGTTATTTAA AATTTGGAT7 4560
CAACAGGGAA GATGGTTGCA TAGTCTATCG TGAATGGGCT CCTGCTGCTC AGTAGGTCC7 4620
CGTCTACTAC AAAATAGTAG TTTCCATCAT CATAACAGAT TTTCCTATTA AAGCATGATG 4680
TTGCAGCATC ATTGGCTTTC 7TACATGTTC TAATTGCTAT TAAGGTTATG CTTCTAATTA 4740
ACTCATCCAC AATGCAGGGA AGCAGAAGTT ATTGsCGATT TCAATGGATG GAACGGTTCT 4800
AACCACATGA TGGAGAAGGA CCAGTTTGGT GTTTsGAGTA TTAGAATTCC TGATGTTGAC 4860
AGTAAGCCAG TCATTCCACA CAACTCCAGA GTTAAGTTTC GTTTCAAACA TGGTAATGGA 4920
GTGTGGGTAG ATCGTATCCC 7GC7TGGATA AAGTATGCCA CTGCAGACGC CACAAAGTTT '4980
GCAGCACCAT ATGATGGTGT CTACTGGGAC CCACCACCTT CAGAAAGGTT TTsTTATTCA 5040
TACCTTGAAG CTGAATTTTG AACACCATCA TCACAGGCAT TTCGATTCAT GTTCTTACTA 5100
GTCTTGTTAT GTAAGACATT TTGAAATGCA AAAG77AAAA TAATTGTGTC T7TACTAA77 5160
OGACTTGAT CCCATACTCT TTCCCTTAAC AAAATGAGTC AATTCTATAA GTGCTTGAGA 5220
ACT7AC7AC77CAGCAA77A AACAGGTACC ACTTCAAATA CCC7CGCCCT CCCAAACCCC 5280
GAGCCCCACG AA7C7ATGAA GCACATGTCG GCATGAGCAG CTCTGAGCCA CGTGTAAA77 5340
CG7A7CG7GA G777GCAGA7 GATGTTTTAC CTCGGATTAA GGCAAATAAC TATAATACTG 5 00
7CCAG77GA7 GGCCATAA7G GAACATTCTT ACTATGGATC ATTTGGATAT CATGTTACAA 5460
ACTTTT77GC 7G7GAGCAG7 AGATATGGAA ACCCGGAGGA CCTAAAGTAT CTGATAGA7A 5520
AAGCACATAG CTTsGGTTTA CAGGTTCTGG TGGATGTAGT TCACAGTCAT GCAAGCAATA 5580
ATGTCACTGA TsGCC7CAA7 GGCTTTGATA TTGGCCAAGG TTCTCAAGAA TCCTACTTTC 5640
ATGCTGGAGA GCGAGGGTAC CATAAGTTGT GGGATAGCAG GCTGTTCAAC TATGCCAATT 5700
GGGAGGTTCT TCG77TCCTT CTTTCCAACT TGAGG7GGTG GCTAGAAGAG TATAACTTTG 5760
ACGGATT7CG A7TTGATGGA ATAACTTCTA TGCTGTATGT TCATCATGGA ATCAATATGG 5820
GATTTACAGG AAACTATAAT GAGTATTTCA GCGAGGCTAC AGATGTTGAT GCTGTGGTCT 5880
ATTTAATGTT GGCCAATAAT CTGATTCACA AGATTTTCCC AGATGCAACT GTTATTGCCG 5940
AAGATGTTTC TGGTATGCCG GGCCTTGGCC GGCCTGTTTC TGAGGGAGGA ATTGGTTTTG 6000
TTTACCGCCT GGCAATGGCA ATCCCAGATA AGTGGATAGA TTATTTAAAG AATAAGAATG S060
A7GAAGA7TG G7CCA7GAAG GAAGTAACAT CGAGTTTGAC AAATAGGAGA TATACAGAGA 6120
AGTGTATAGC ATATGCGGAG ACCCATGATC AGGTATTT7A AATTTATTTC TACAACTAAA 6180
TAATTCTCAG AACAATTGTT AGATAGAATC CAAATATATA CGTCCTGAAA GTATAAAAG7 6240
ACTTATTTTC GCCATGGGCC TTCAGAATAT TGGTAGCCGC TGAATATCAT GATAAGTTAT 6300
TTATCCAGTG ACATTTTTAT GTTCACTCCT ATTATGTCTG CTGGATACAG tCTATTGTTG 6360
GTGACAAGAC CATTGCATTT CTCCTAATGG ACAAAGAGAT GTATTCTGGC ATGTCTTGCT 6420
TGACAGATGC 7TCTCCTGTT GTTGATCGAG GAATTGCGCT TCACAAGGTT TGTCTGTTTC 6480
TATTGCATTT TAAGGTTCAT ATAGGTTAGC CACGGAAAAT CTCACTCTTT GTGAGGTAAC 6540
CAGGGTTCTG ATGGATTATT CAATTTTCTC GTTTATCATT TGTTTATTCT TTTCATGCAT 6600
TGTGTTTCTT TTTCAATATC CCTCTTATTT GGAGGTAATT TTTCTCATCT ATTCACTTTT 6660CACAGATGAT CCATTTTTTC ACAATGGCCT TGGGAGGAGA GGGGTACCTC 6720
AATTTCATGG GTAACGAGGT ATGTCTTACA TCTTTAGATA TTTTGTGATA ATTACAATTA 6780
GTTTGGCTTA CTTGAACAAG ATTCATTCCT CAAAATGACC TGAACTGTTG AACATCAAAG 6840
GGGTTGAAAC ATAGAGGAAA ACAACATGAT GAATGTTTCC ATTGTCTAGG GA TTCTATT 6900 ATGTTGCTGA GAACAAATGT CATCTTAAAA AAAACATTGT TTACTTTTTT GTAGTATAGA 6960
AGATTACTGT ATAGAGTTTG CAAGTGTGTC TGTTTTGGAG TAATTGTGAA ATGTTTGATG 7020
AACTTGTACA GTTTGGCCAT CCTGAGTGGA TTGACTTCCC TAGAGAGGGC AATAATTGGA 7080
GTTATGACAA ATGTAGACGC CAG7GGAACC 7CGCGGATAG CGAACACTTG AGATACAAGG 7140
TTCAAGTATT TTGAATCGCA GCTTGTTAAA TAATCTAGTA ATTTTTAGAT TGCTTACTTG 7200
GAAGTCTACT TGGTTCTGGG GATGATAGCT CATTTCATCT TGTTCTACTT ATTTTCCAAC 7260
CGAATTTCTG ATTTTTTTTTT CGAGATCCAA GTATTAGATT CATTTACACT TATTACCGCC 7320
TCATTTCTAC CACTAAGGCC TTGATGAGCA GCTTAAGTTG ATTCTTTGAA GCTATAGTTT 7380
CAGGCTACCA ATCCACAGCC TGCTATATTT GTTGGATACT TACCTTTTCT TTACAATGAA 7440
GTGATACTAA TTGAAATGGT CTAAATCTGA TATCTATATT TCTCCGTCTT TCCTCCCCCT 7500
CATGATGAAA TGCAGTTTAT GAATGCATTT GATAGAGCTA TGAATTCGCT CGATGAAAAG 7560
TTCTCATTCC TCGCATCAGG AAAACAGATA GTAAGCAGCA TGGATGATGA TAATAAGGTA 7620
AAATCATCTA AAGTTGAAAG TGTTGGGTTT ATGAAGTGCT TTAATTCTAT CCAAGGACAA 76B0
GTAGAAACCT TTTTACCTTC CATTTCTTGA TGATGGATTT CATATTATTT AATCCAATAG_7740_CTGGTCAAAT TCGGTAATAG CTGTACTGAT TAGTTACTTC ACTTTGCAGG TTGTTGTGTT 7800
TGJAACGTGGT GACCTGGTAT TTGTATTCAA CTTCCACCCA AAGAACACAT ACGAAGGGTA 7860
TATATGTTTT ACTTATCCAT GAAATTATTG CTCTGCTTGT TTTTAATGTA CTGAACAAGT 7920
TTTATGGAGA AGTAACTGAA ACAAATCATT TTCACATTGT CTAATTTAAC TCTTTTTTTCT 7980
GATCCTCGCA TGACGAAAAC AGGTATAAAG 7TGGATGTGA CTTGCCAGGG AAGTACAGAG 8040
TTGCACTGGA CAGTGATGCT 7GGGAAT77G GTGGCCATGG AAGAGTAAGG ATTTGCT7GA 8100
ATAAC7777G A7AA7AAGA7 AACAGA7G7A GGG7ACAGTT CTCTCACCAA AAAGAACTGT 8160
AATTGTCTCA TCCATC7TTA G77GTATAAG ATATCCGACT GTCTGAGTTC GGAAGTGTT7 8220
GAGCC7CCTG CCCTCCCCCT GCGTTGTTTA GCTAAT7CAA AAAGGAGAAA ACTGTTTA7T 8280
GATGATCTTT GTCTTCA7GC 73ACATACAA TCTGTTCTCA TGACAGACTG GTCATGATGT 8340
TGACCATTTC ACATCACCAG AAGGAATACC TGGAGTTCCA GAAACAAATT TCAATGGTCG 8400
TCCAAA7TCC T7CAAAG7GC 7GTCTCCTGC GCGAACATGT GTGGTACAGT TCTTGCCGTG 8460
TGACC7CCC7 T7TTAT7G7G G7T7TG7TCA 7AGTTATTTG AATGCGATAG AAGT7AAC7A 8520 tTGATTACCG CCACAATCGC CAGTTAAG7C CTCTGAACTA CTAATTTGAA AGGTAGGAAT 8580
AGCCGTAATA AGG7C7AC77 77GGCATCTT ACTGTTACAA AACAAAAGGA TGCCAAAAAA 8640
ATTCTTCTCT ATCC7C7777 7CCC7AAACC AGTGCATGTA GCTTGCACCT GCATAAAC77 8700
AGGTAAATGA TCAAAAA7GA AGTTGATGGG AACTTAAAAC CGCCCTGAAG TAAAGCTAGG 8760
AATAGTCATA TAATGTCCAC CTTTGGTGTC TGCGC7AACA TCAACAACAA CATACCTCGT 8820
GTAGTCCCAC AAAGTGG777 CAGGGGGAGG GTAGAGTGTA TGCAAAACTT ACTCCTATCT 8880
CAGAGGTAGA GAGGAT7TTT TCAATAGACC CTTGGCTCAA GAAAAAAAGT CCAAAAAGAA 8940
GTAACAGAAG TGAAAGCAAC ATGTGTAGCT AAAGCGACCC AACTTGTTTG GGACTGAAGT 9000
AGTTsTTGTT GTTGAAACAG TGCATGTAGA TGAACACATG TCAGAAAATG GACAACACAG 9060
TTATTTTGTs CAAGTCAAAA AAATGTACTA CTATTTCTTT GTGCAGCTTT ATGTATAGAA 9120
AAGTTAAATA ACTAATGAAT 7TTGCTAGCA GAAAAATAGC TTGGAGAGAA ATTTTTTATA 9180
TTGAACTAAG CTAACTATAT TCATCTTTCT TTTTGCTTCT TCTTCTCCTT GTTTGTGAAG 9240
GCTTATTACA GAGTTGATGA ACGCATGTCA GAAACTGAAG ATTACCAGAC AGACATTTGT 9300
AGTGAGCTAC TACCAACAGC CAATATCGAG GAGAGTGACG AGAAACTTAA AGATTCGTTA 9360
TCTACAAATA TCAGTAACAT TGACGAACGC ATGTCAGAAA CTGAAGTTTA CCAGACAGAC 9420 ATTTCTAGTG AGCTACTACC AACAGCCAAT ATTGAGGAGA GTGACGAGAA ACTTAAAGA7 9480
TCGTTATCTA CAAATATCAG 7AACA7TGAT CAGACTGTTG TAGTTTCTGT TGAGGAGAGA 9540
GACAAGGAAC 7TAAAGAT7C ACCG7C7G7A AGCATCATTA GTGATGTTGT TCCAGCTGAA 9600
TGGGATGATT CAGATGCAAA CGTCTGGGGT GAGGACTAGT CAGATGATTG ATCGACCCT7 9660
CTACCGATTG GTGATCGCTA TCC7TGCTC7 CTGAGAAATA GGTGAGGCGA AACAAAAAA7 9720
AATTTGCATG ATAAAAAGTC TGATTTTATG ATCGCTATCC TCGCTCTCTG AGAAAGAAGC 9780
GAAACAAAGG CGACTCCTGG ACTCGAATCT ATAAGATAAC AAAGGCGACT CCTGGGACTC 9840 GAATCTATAA GATAACAAAG GCAATTCCAA GACTTGAATC TATAAAAAAT TTAGTTAAGA 9900
ATGATTAACG TCCGATCCTA ATTCGAATCG AGGCATCTTA CCACTCCATT GATAATTATA 9960
TAAGTCAATA AGTCATATAA WAGTATTAAA AACTAAATTG ACTTGATCGG TCTATCAAAA 10020
ATMAGATMAA ATTGTGTTCA TATGTAACAT TTTTGTTGTC ACAATTAGCT TAATTACATC 10080
TTTCATGTGC AATAACAAAG AAATGATAGG AATTTAGAGA TTCCAATTTT TTTGTTGCCA 10140
CAATTAACTT AATTACATCT 7TCATTTGCA ATAACAAAGA AATGATAGGA ATTTAGAGAT 10200
CCAGTGTCAA TACACAACCT AGGCCAACAT CGAAAGCATA ACTGTAAACT CATGCATGAA 10260
GAAATCAGTC GTAAAAATGA ATAAATGCGA CATAAAAACA AATTGCATGT ATCATTAATG 10320 TGACTTAACT ACAAGTAAAA AtAAATTTAA CAAATGTAAC TTSACTACAA GTAAAAATAA 10380
ATTGCTTCTA 7CATTAACAA ACAAACAGAA TTAAAAAGAA AAAAACATAC TAAA7C77AC 10440
CGTCATTCGA TAAAAAAAAA 7ACCAAATTC ATAATGCAAG GAAAACGAAA CGCGTCCTGA 10500
TCGGGTATCA ACGATGAAAT GGACCAGTTG GATCGACTGC CTGCACAACG TTAGGTATGC 10560
CAAAAAAAAG AACACGATCC TTTGCACCCG TTCGATGATT ATCAGTATGT TCACAAAAAA 10620
AACTTAAGTT CATCCCAGTG TACAACAGCC CCAACATCTG CCCCAAGTAA CAAAAAACAA 10680
CCAATTTATC TTATTCTTAT CTGCCACAAA ATAATCGGTT TCACACTATT CTCTTGTTAT 10740
ACAAAATTGA CAAGTAGGAA GGAGAGGAGT CATCCAAATA AACGGTGCAC GTTCTTTGAG 10800 AAAAGTCTTA T7TTTCGTAA GATCCAATTT CAACAAACTT TTCTTCAAGT CAAAATTCCT 10860
GATAGTGTAT C7CCTCTCGA CGACC7CTTG CATTGAACGA TCTCCGCTTA TCATGAAAAG 10920
TTGCTTGGAT AACAAGTATT GCAAGGGGGG GACAGTAGCT ATTAAGTTAG TCGGCCCAAG 10980
GAAATGGAGG AGTGATAGTC 7CGAA7A77A 7TCACCTCTT TAGCATTACC CGGTCTGGCT 11040
TTAAGGAGTT ACGTCTTTTA CGCTCGCCAA T'n'CT'l ri TAGAATGGTT GGTGTCAAAA 11100
TCGCGAGTTG TGGAAGGTTC AAGTTACTCG ATTCGTGATT TTCAAGTATG AGTGGTGAGA 11160
GAGATTCGAT ATTTTCACGA GGTGTATTCG AGGTCTAGTA GAACGAAGGG TGTCACTAAT 11220
GAAAGTTTCA AGAGTTCATC ATCATCTTCT TCTAGTAGAT TTTCGCTTTC AAATGAGTAT 11280 GAAAATTCTT CCTCTTTTCT ATTGATTTTC TTCATTGTTT TCTTCATTGT TGTGGtTGTT 11340
ATTGAAAAGA AAGAAAA777 ATAACAGAAA AAGATGTCAA AAAAAAGGTA AAATGAAAGA 11400
GTATCATATA CTTAAAGAGT TGCGTAGAGA TAAGTCAAAA GAAACAGAAT TATAGTAATT 11460
TCAGCTAAGT TAGAATTC 11478
Claims (13)
1. A method for carrying out the enzymatic activity in a plant (or a cell, a tissue or an organ thereof) which comprises expressing in the plan + a (a cell a tissue or an organ thereof) a nucleus + sequence 1 two also characterized because the nucleus sequence + gone is encoded, partially or completely, pair-to an index + in a sense orientation; and by consequence of nucleotides it does not contain a sequence that is in sense to a sequence of exons normally associated with the tron.
2. A method of compliance with claim 1, further characterized in that the activity of the starch enrichment enzyme is affected and / or that the levels of the arylopectm are affected and / or the composition of the starch is changed.
3. A method to affect the enzymatic activity in an organism that produces starch (a cell, a tissue or an organ itself) that comprises expressing in the organism (or a cell, a + e? Do or an organ thereof) which produces a sequence of nucleotides, further characterized in that the nucleotide sequence is encoded, partially or completely, for an intron in a seated axis orientation; where the nucleotide sequence does not contain a sequence that is sense for a sequence exon normally associated with m +? * on; and because the activity of the starch enzyme is affected and / or the levels of the protein are changed, and / or the composition of the starch is changed.
4. A method according to any of claims 1 to 3, further characterized in that the nucleotide sequence does not contain a sequence that is in sense to a sequence of exons normally associated with the intron.
5. A method according to any of the preceding claims, further characterized in that the enzymatic activity is reduced or eliminated.
6. A method according to any of the preceding claims, further characterized in that the nucleotide sequence is encoded at least substantially by at least a whole host in a sense orientation.
7. A method according to any of the preceding claims, further characterized in that the nucleotide sequence is coded for a whole mtron in a sense orientation.
8. A method according to any of the preceding claims, further characterized in that the sequence of nucleotides comprises the sequence shown as any of TD. SEC. No. 1 to ID. SEC. No. 13 or a variant, a derivative or a homologous thereof, including combinations thereof.
9. - A method according to any of the preceding claims, further characterized in that the nucleotide sequence is expressed by a promoter having a sequence shown as TD. SEC. No. 14 or a variant, a derivative or a homologous thereof.
10. A sequence in the sense comprising the nucleotide sequence defined in claim 8 or a vanant, a derivative or a homologous thereof. 11.- A promoter that has a sequence shown as TD. SEC. No. 14, or a variant, a derivative, a counterpart thereof. 1 .- A promoter * according to claim 11 in combination with? N gene of interest ("GOI"). 13. A construction capable of understanding or expressing the invention in accordance with any of claims 10 to 12. 14.- A vector- comprising or expressing the invention in accordance with any of claims 10 to 13. 15.- A combination of nucleotide sequences comprising a first nucleotide sequence that is encoded for a recombinant enzyme; and a second nucleotide sequence corresponding to a món in sense orientation; because the intron is a mtron that is associated with a genomic gene that encodes an enzyme corresponding to a recornbinant enzyme; and because the second nucleotide sequence does not contain a sequence that is in the sense of a sequence of exons normally associated with the intron. 16.- A cell, a tissue or an organ comprising or expressing the invention according to any one of claims LO to 15. 17. A transgenic starch-producing organism comprising and expressing the invention in accordance with any of claims 10 to 16. .- A transgenic producer of starch according to application 17, also characterized because the organism is a plant. 19. A starch obtained from the invention according to any of the preceding claims. 20.- pBEAll (NCIMB 40754). 21. A nucleotide sequence that is antisense to any or all of the int sequence sequences obtainable from? -SBE 3.2 (NCTMB 40751) or? - SBE 3.4 (NCTMB 40752) or a variant, a derivative or a homologous of them. 22. A method for expressing a protein or reconstituting enzyme in a host organism comprising expressing a nucleotide sequence that is encoded for a protein or a reclosing enzyme; and expressing an additional nucleotide sequence, further characterized in that the additional nucleotide sequence is encoded, partially or completely, for a motor in a sense orientation; because the intron is an intron normally associated with the genomic gene that encodes a protein or an enzyme corresponding to the protein or recom- binant enzyme; and because the additional nucleotide sequence does not contain a sequence that is sended to a sequence of exons normally associated with the intron
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9514437.4 | 1995-07-14 | ||
| GBGB9514437.4A GB9514437D0 (en) | 1995-07-14 | 1995-07-14 | Inhibition of gene expression |
| PCT/EP1996/003053 WO1997004113A2 (en) | 1995-07-14 | 1996-07-12 | Inhibition of gene expression |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| MXPA98000499A true MXPA98000499A (en) | 1998-04-01 |
| MX9800499A MX9800499A (en) | 1998-04-30 |
Family
ID=10777674
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| MX9800499A MX9800499A (en) | 1995-07-14 | 1996-07-12 | Inhibition of gene expression. |
Country Status (11)
| Country | Link |
|---|---|
| US (1) | US6232122B1 (en) |
| EP (1) | EP0839203A1 (en) |
| JP (1) | JPH11509413A (en) |
| CN (1) | CN1196092A (en) |
| AU (1) | AU707758B2 (en) |
| BR (1) | BR9609797A (en) |
| CA (1) | CA2226994A1 (en) |
| GB (1) | GB9514437D0 (en) |
| MX (1) | MX9800499A (en) |
| NZ (1) | NZ313627A (en) |
| WO (1) | WO1997004113A2 (en) |
Families Citing this family (28)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ336607A (en) * | 1997-02-21 | 2001-06-29 | Danisco | Method of inhibiting gene expression where the nucleotide codes for a Class A SBE intron in the sense direction |
| CA2280230A1 (en) | 1997-02-21 | 1998-08-27 | Danisco A/S | Antisense intron inhibition of starch branching enzyme expression |
| GB9706381D0 (en) * | 1997-03-27 | 1997-05-14 | Cambridge Advanced Tech | Improvements relating to the specificity of gene expression |
| GB9709161D0 (en) * | 1997-05-06 | 1997-06-25 | Danisco | A process of preparing an anti-oxidant |
| ES2321248T3 (en) | 1997-09-12 | 2009-06-03 | Commonwealth Scientific And Industrial Research Organisation | REGULATION OF GENE EXPRESSION IN PLANTS. |
| AU727294B2 (en) * | 1997-09-12 | 2000-12-07 | Biogemma Sas | Regulation of gene expression in plants |
| US6426185B1 (en) | 1998-01-16 | 2002-07-30 | Large Scale Biology Corporation | Method of compiling a functional gene profile in a plant by transfecting a nucleic acid sequence of a donor plant into a different host plant in an anti-sense orientation |
| US6303848B1 (en) | 1998-01-16 | 2001-10-16 | Large Scale Biology Corporation | Method for conferring herbicide, pest, or disease resistance in plant hosts |
| US20060005282A1 (en) * | 2002-04-23 | 2006-01-05 | Activx Biosciences, Inc | Production and use of salt tolerant and culture density tolerant organisms |
| EP2216405A1 (en) | 2002-05-03 | 2010-08-11 | Monsanto Technology LLC | Speed specific USP promoters for expressing genes in plants |
| US7078234B2 (en) | 2002-12-18 | 2006-07-18 | Monsanto Technology Llc | Maize embryo-specific promoter compositions and methods for use thereof |
| DK1578973T3 (en) | 2002-12-19 | 2008-11-24 | Bayer Cropscience Ag | Plant cells and plants that synthesize a starch with increased final viscosity |
| EP1613730A4 (en) | 2003-03-28 | 2007-12-05 | Monsanto Technology Llc | NEW PLANT PROMOTERS FOR USE IN EARLY DEVELOPMENT OF SEEDS |
| EP1649022B1 (en) | 2003-06-30 | 2011-07-27 | Commonwealth Scientific And Industrial Research Organisation | Wheat with altered branching enzyme activity and starch and starch containing products derived thereform |
| AU2006217847B2 (en) | 2005-02-26 | 2011-04-07 | Basf Plant Science Gmbh | Expression cassettes for seed-preferential expression in plants |
| EP1707632A1 (en) | 2005-04-01 | 2006-10-04 | Bayer CropScience GmbH | Phosphorylated waxy potato starch |
| CN101198701B (en) * | 2005-04-19 | 2013-04-24 | 巴斯福植物科学有限公司 | Endosperm-specific and/or germinating embryo-specific expression of starch in monocots |
| EP1874937A2 (en) | 2005-04-20 | 2008-01-09 | BASF Plant Science GmbH | Expression cassettes for seed-preferential expression in plants |
| BRPI0614070A2 (en) | 2005-05-10 | 2018-07-31 | Basf Plant Science Gmbh | expression cassettes, isolated nucleotide sequence, synthetic transcriptional regulatory sequence, methods for providing a synthetic transcriptional regulatory nucleotide sequence, and an expression cassette, vector, non-human organism or transgenic host cell (a), and, cell of plant or transgenic plant |
| CN101631868B (en) | 2007-02-16 | 2016-02-10 | 巴斯福植物科学有限公司 | For regulating the nucleotide sequence of embryo-specific expression in monocotyledons |
| US8799004B2 (en) * | 2007-07-05 | 2014-08-05 | Map Ip One, Llc | System and method for real estate spatial data analysis |
| WO2010118477A1 (en) | 2009-04-17 | 2010-10-21 | Molecular Plant Breeding Nominees Ltd | Plant promoter operable in endosperm and uses thereof |
| CA2766858A1 (en) | 2009-07-10 | 2011-01-13 | Basf Plant Science Company Gmbh | Expression cassettes for endosperm-specific expression in plants |
| AU2010325720A1 (en) | 2009-12-03 | 2012-07-19 | Basf Plant Science Company Gmbh | Expression cassettes for embryo-specific expression in plants |
| BR112014008723A2 (en) | 2011-10-12 | 2019-09-24 | Bayer Cropscience Ag | plants with decreased activity of a starch dephorylation enzyme |
| BR112014008895A2 (en) | 2011-10-12 | 2019-09-24 | Bayer Cropscience Ag | plants with decreased activity of a starch dephosphorylation enzyme |
| HUE067217T2 (en) | 2016-05-26 | 2024-10-28 | Nunhems Bv | Seedless fruit producing plants |
| WO2019238832A1 (en) | 2018-06-15 | 2019-12-19 | Nunhems B.V. | Seedless watermelon plants comprising modifications in an abc transporter gene |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4740463A (en) * | 1984-04-13 | 1988-04-26 | Massachusetts Institute Of Technology | Methods and artificial genes for antagonizing the function of an oncogene |
| US5034323A (en) * | 1989-03-30 | 1991-07-23 | Dna Plant Technology Corporation | Genetic engineering of novel plant phenotypes |
| SE9004095L (en) * | 1990-12-21 | 1992-06-01 | Amylogene Hb | GENETIC MODIFICATION OF POTATOIS BEFORE EDUCATION OF AMYLOST TYPE |
| NZ241310A (en) * | 1991-01-17 | 1995-03-28 | Gen Hospital Corp | Trans-splicing ribozymes |
| DE4104782B4 (en) * | 1991-02-13 | 2006-05-11 | Bayer Cropscience Gmbh | Novel plasmids containing DNA sequences that cause changes in carbohydrate concentration and carbohydrate composition in plants, as well as plants and plant cells containing these plasmids |
| AU663702B2 (en) * | 1991-03-06 | 1995-10-19 | Board Of Regents, The University Of Texas System | Methods and compositions for the selective inhibition of gene expression |
| JPH0698656A (en) * | 1992-03-30 | 1994-04-12 | Mitsui Giyousai Shokubutsu Bio Kenkyusho:Kk | Rice plant treated with gene |
| DE69233352T2 (en) * | 1992-10-14 | 2004-10-07 | Syngenta Ltd | PLANTS AND METHOD FOR THEIR PRODUCTION |
| GB9223454D0 (en) * | 1992-11-09 | 1992-12-23 | Ici Plc | Novel plants and processes for obtaining them |
| CA2186399C (en) * | 1994-03-25 | 2001-09-04 | David Cooke | Method for producing altered starch from potato plants |
-
1995
- 1995-07-14 GB GBGB9514437.4A patent/GB9514437D0/en active Pending
-
1996
- 1996-07-12 CN CN96196982A patent/CN1196092A/en active Pending
- 1996-07-12 MX MX9800499A patent/MX9800499A/en unknown
- 1996-07-12 CA CA002226994A patent/CA2226994A1/en not_active Abandoned
- 1996-07-12 AU AU66146/96A patent/AU707758B2/en not_active Ceased
- 1996-07-12 EP EP96925732A patent/EP0839203A1/en not_active Withdrawn
- 1996-07-12 BR BR9609797A patent/BR9609797A/en not_active Application Discontinuation
- 1996-07-12 NZ NZ313627A patent/NZ313627A/en unknown
- 1996-07-12 US US08/983,440 patent/US6232122B1/en not_active Expired - Fee Related
- 1996-07-12 JP JP9506261A patent/JPH11509413A/en active Pending
- 1996-07-12 WO PCT/EP1996/003053 patent/WO1997004113A2/en not_active Ceased
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| MXPA98000499A (en) | Inhibition of the expression of ge | |
| AU707758B2 (en) | Inhibition of gene expression | |
| AU709798B2 (en) | Inhibition of gene expression | |
| US6483009B1 (en) | Antisense intron inhibition of starch branching enzyme expression | |
| AP1169A (en) | Regulation of quinolate phosphoribosyl transferase expression. | |
| JP2993979B2 (en) | Method for producing transgenic tuber-forming dicotyledonous plant, plant cell containing expression cassette, patatin promoter sequence and expression cassette | |
| MXPA98000500A (en) | Inhibition of the expression of ge | |
| US6444878B1 (en) | Method of plant selection using glucosamine-6-phosphate deaminase | |
| CA2280210A1 (en) | Sense intron inhibition of starch branching enzyme expression | |
| US5723757A (en) | Plant promoters specific for sink organ expression of genes | |
| WO1998054335A1 (en) | In vivo modification of galactomannans in guar by expression of udp-galactose epimerase antisense rna | |
| US6025544A (en) | Processes for modifying plant flowering behavior | |
| JPH10512451A (en) | Deoxyribonucleic acid encoding glutathione S-transferase and use thereof | |
| AU2004203879B2 (en) | Regulation of quinolate phosphoribosyl transferase expression | |
| AU5189399A (en) | Selection method | |
| AU2002300895B2 (en) | Regulation of quinolate phosphoribosyl transferase expression | |
| MXPA99007795A (en) | Sense intron inhibition of starch branching enzyme expression | |
| MXPA99007348A (en) | Selection method for transgenic plants | |
| MXPA99007796A (en) | Antisense intron inhibition of starch branching enzyme expression |