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ES2594284T3 - Variantes de HPPD y procedimientos de uso - Google Patents

Variantes de HPPD y procedimientos de uso Download PDF

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Publication number
ES2594284T3
ES2594284T3 ES13164699.4T ES13164699T ES2594284T3 ES 2594284 T3 ES2594284 T3 ES 2594284T3 ES 13164699 T ES13164699 T ES 13164699T ES 2594284 T3 ES2594284 T3 ES 2594284T3
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pos
amino acid
hppd
twenty
thaliana
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Gudrun Lange
Fabien Poree
Bernd Laber
Jörg Freigang
Arno Schulz
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Bayer CropScience AG
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Peptides Or Proteins (AREA)
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Abstract

Un ácido nucleico aislado que comprende una secuencia de nucleótidos que codifica una proteína de HPPD mutada, en el que dicha proteína de HPPD mutada tiene actividad de HPPD y en el que en dicha proteína de HPPD mutada el aminoácido en una posición correspondiente a la posición 252 de la SEC ID N.º: 2 ha sido reemplazado de tal modo que la secuencia de aminoácidos resultante comprende un aminoácido seleccionado de His, Ile, Leu, Asn o Ser en la posición correspondiente a la posición 252 de la secuencia de aminoácidos de la SEC ID N.º: 2:

Description

imagen1
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Tabla 1: aminoácidos que forman el sitio de unión en A. thaliana, P. fluorescens, S. avermitilis, H. sapiens, R. norvegicus [no formando parte de la invención]
Arabidopsis thaliana
Pseudomonas fluorescens Streptomyces avermitilis Homo sapiens Rattus norvegicus
Posición Aminoácido
Posición Aminoácido
Posición Aminoácido
Posición Aminoácido
Posición Aminoácido
226 H 228 V 248 H 250 F 251 A
162 H 164 T 186 R 188 A 189 R 187 H 189 V 211 T 213 M 214 K 183 H 185 V 207 H 209 F 210 W 183 H 185 V 207 H 209 F 210 W
252 E 253 F 265 L 267 S 268 A 269 V 270 L 271 A 280 P 282 N 293 Q 294 I 307 Q 308 H 335 M 342 Y 368 L 379 Q 381 F 392 F 394 E 419 F 420 G 421 K 422 G 423 N 424 F 425 S 426 E 427 L 431 I
190 Y 191 F 200 L 202 S 203 K 204 A 205 M 206 S 215 P 217 N 226 Q 227 I 240 Q 241 H 264 M 271 Y 295 L 310 Q 312 F 321 F 323 E 333 F 334 G 335 E 336 G 337 N 338 F 339 K 340 A 341 L 345 I 215 E 216 F 228 L 230 S 231 K 232 V 233 V 234 A 243 P 245 N 255 Q 256 I 269 Q 270 H 293 L 299 Y 323 L 334 Q 336 F 347 F 349 E 359 F 360 G 361 K 362 G 363 N 364 F 365 K 366 A 367 L 371 I 211 S 212 V 224 L 226 S 227 I 228 V 229 V 230 A 239 P 241 N 251 Q 252 I 265 Q 266 H 289 L 295 Y 323 L 334 Q 336 F 347 F 349 E 359 F 360 G 361 A 362 G 363 N 364 F 365 N 366 S 367 L 371 F 211 S 212 V 224 L 226 S 227 I 228 V 229 V 230 A 239 P 241 N 251 Q 252 I 265 Q 266 H 289 L 295 Y 323 L 334 Q 336 F 347 F 349 E 359 F 360 G 361 A 362 G 363 N 364 F 365 N 366 S 367 L 371 F
Una alineación de ejemplo de las proteínas de HPPD se da en la Tabla 2a para las proteínas de HPPD con estructuras 3D conocidas. La Tabla 2a da la numeración de los aminoácidos de la secuencia de Arabidopsis y también los aminoácidos que son comunes dentro de estas secuencias de HPPD, designando estos aminoácidos con un asterisco. Sobre la base de tal alineación y a partir de la definición del aminoácido de Arabidopsis por su posición y su naturaleza, es fácil identificar la posición del correspondiente aminoácido en otra secuencia de HPPD.
8
La Figura 2 muestra que esto se puede llevar a cabo con la alineación de las secuencias de diferente origen vegetal, mamífero y bacteriano, lo que demuestra que este procedimiento de alineación, que es bien conocido por el experto en la técnica, puede ser generalizado a cualquier otra secuencia. Una alineación de diferentes secuencias de HPPD también se describe en la solicitud de patente WO 97/49816.
Tabla 2a: alineación de secuencias de HPPD con estructuras cristalinas conocidas, es decir, A. thaliana, P. fluorescens, S. avermitilis, H. sapiens, R. norvegicus [no formando parte de la invención]
Pos. 1TFZ Pos. 1CJX Pos. 1T47 Pos. 3ISQ Pos. 1SQI Común
A. thaliana P. fluorescens S. avermitilis H. sapiens R. norvegicus aminoácido
1M--2G--3H --4Q--5N --6A --7A --8V --9S --
10E --11N --12Q--13N --14H -1M 15D -2T 16D -3Q 17G -4T 18A -5T 19 A -6H8G8G 20 S -7H9A9P 21 S -8T10K10K 22 P -9P11P 11P 23G--24F --25K --26L --27V --28G--29F --30S --31K --32F --33V --34 R -10D12E 12E 35 K 2A 11T13R 13R
9
(continuación)
Pos.
1TFZ Pos. 1CJX Pos. 1T47 Pos. 3ISQ Pos. 1SQI Común
A. thaliana
P. fluorescens S. avermitilis H. sapiens R. norvegicus aminoácido
36
N 3 D 12 A 14 G 14 G
37
P 4 L 13 R 15 R 15 R
38
K 5 Y 14 Q 16 F 16 F
39
S 6 E 15 A 17 L 17 L
40
D 7 N 16 D 18 H 18 H
41
K 8 P 17 P -
42
F 9 M 18 F -
43
K 10 G 19 P -
44
V 11 L 20 V -
45
K 12 M 21 K -
46
R 13 G 22 G -
47
F 14 F 23 M 19 F 19 F
48
H 15 E 24 D 20 H 20 H
49
H 16 F 25 A 21 S 21 S
50
I 17 I 26 V 22 V 22 V
51
E 18 E 27 V 23 T 23 T
52
F 19 F 28 F 24 F 24 F x
53
W 20 A 29 A 25 W 25 W
54
C 21 S 30 V 26 V 26 V
55
G 22 P 31 G 27 G 27 G
56
D 23 T 32 N 28 N 28 N
57
A 24 P 33 A 29 A 29 A
58
T 25 G 34 K 30 K 30 K
59
N 26 T 35 Q 31 Q 31 Q
60
V 27 L 36 A 32 A 32 A
61
A 28 E 37 A 33 A 33 A
62
R 29 P 38 H 34 S 34 S
63
R 30 I 39 Y 35 F 35 F
64
F 31 F 40 Y 36 Y 36 Y
65
S 32 E 41 S 37 C 37 C
66
W 33 I 42 T 38 S 38 N
67
G 34 M 43 A 39 K 39 K
68
L 35 G 44 F 40 M 40 M
69
G 36 F 45 G 41 G 41 G
70
M 37 T 46 M 42 F 42 F
71
R 38 K 47 Q 43 E 43 E
72
F 39 V 48 L 44 P 44 P
73
S 40 A 49 V 45 L 45 L
74
A 41 T 50 A 46 A 46 A
75
K 42 H 51 Y 47 Y 47 Y
76
S 43 R 52 S 48 R 48 K
10
(continuación)
Pos.
1TFZ Pos. 1CJX Pos. 1T47 Pos. 3ISQ Pos. 1SQI Común
A. thaliana
P. fluorescens S. avermitilis H. sapiens R. norvegicus aminoácido
77
D 44 S 53 G 49 G 49 G
78
L 45 K 54 P 50 L 50 L
79
S 46 N 55 E 51 E 51 E
80
T - 56 N 52 T 52 T
81
G - 57 G 53 G 53 G
82
N - 58 S 54 S 54 S
83
M - 59 R 55 R 55 R
84
V - 60 E 56 E 56 E
85
H - 61 T 57 V 57 V
86
A - 62 A 58 V 58 V
87
S 47 V 63 S 59 S 59 S
88
Y 48 H 64 Y 60 H 60 H
89
L 49 L 65 V 61 V 61 V
90
L 50 Y 66 L 62 I 62 I
91
T 51 R 67 T 63 K 63 K
92
S 52 Q 68 N 64 Q 64 Q
93
G 53 G 69 G 65 G 65 G X
94
D 54 E 70 S 66 K 66 K
95
L 55 I 71 A 67 I 67 I
96
R 56 N 72 R 68 V 68 V
97
F 57 L 73 F 69 F 69 F
98
L 58 I 74 V 70 V 70 V
99
F 59 L 75 L 71 L 71 L
100
T 60 N 76 T 72 S 72 C
101
A 61 N 77 S 73 S 73 S
102
P 62 E 78 V 74 A 74 A
103
Y 63 P 79 I 75 L 75 L
104
S 64 N 80 K 76 N 76 N
105
P 65 S 81 P -
106
S 66 I 82 A -
107
L 67 A 83 T -
108
S 68 S 84 P 77 P 77 P
109
A - 85 W 78 W 78 W
110
G - 86 G 79 N 79 N
111
E - 87 H 80 K 80 K
112
I - 88 F 81 E 81 E
113
K - 89 L 82 M 82 M
114
P - 90 A 83 G 83 G
115
T - - -
116
T - - -
117
T - - -
11
imagen7
imagen8
(continuación)
Pos.
1TFZ Pos. 1CJX Pos. 1T47 Pos. 3ISQ Pos. 1SQI Común
A. thaliana
P. fluorescens S. avermitilis H. sapiens R. norvegicus aminoácido
199
K 135 S 160 D 153 I 153 T
200
S 136 S 161 G 154 G 154 G
201
E 137 I 162 P 155 Q 155 R
202
F 138 Y 163 Y 156 F 156 F
203
L 139 D 164 L 157 L 157 L
204
P 140 I 165 P 158 P 158 P
205
G 141 D 166 G 159 G 159 G
206
F 142 F 167 Y 160 Y 160 F
207
E 143 V 168 V 161 E 161 E
208
R 144 Y 169 A 162 A 162 A
209
V 145 L 170 A 163 P 163 P
210
E 146 E 171 A 164 A 164 T
211
D 147 G - 165 F 165 Y
-
-
-
166 M 166 K
-
-
-
167 D 167 D
-
-
172 P 168 P 168 T
212
A 148 V 173 I 169 L 169 L
213
S 149 E 174 V 170 L 170 L
214
S 150 R 175 E 171 P 171 P
215
F 151 N 176 P 172 K 172 K
216
P 152 P 177 P 173 L 173 L
217
L 153 V 178 A 174 P 174 P
218
D 154 G 179 H 175 K 175 S
219
Y 155 A 180 R 176 C 176 C
220
G 156 G 181 T 177 S 177 N
221
I 157 L 182 F 178 L 178 L
222
R 158 K 183 Q 179 E 179 E
223
R 159 V 184 A 180 M 180 I
224
L 160 I 185 I 181 I 181 I
225
D 161 D 186 D 182 D 182 D x
226
H 162 H 187 H 183 H 183 H x
227
A 163 L 188 C 184 I 184 I
228
V 164 T 189 V 185 V 185 V
229
G 165 H 190 G 186 G 186 G
230
N 166 N 191 N 187 N 187 N x
231
V 167 V 192 V 188 Q 188 Q
232
P 168 Y 193 E 189 P 189 P
-
169 R 194 L 190 D 190 D
-
170 G 195 G 191 Q 191 Q
233
E 171 R 196 R 192 E 192 E
234
L 172 M 197 M 193 M 193 M
14
imagen9
(continuación)
Pos.
1TFZ Pos. 1CJX Pos. 1T47 Pos. 3ISQ Pos. 1SQI Común
A. thaliana
P. fluorescens S. avermitilis H. sapiens R. norvegicus aminoácido
276
M 211 M 239 K 235 S 235 S
277
V 212 I 240 V 236 I 236 I
278
L 213 R 241 K 237 K 237 K
279
L 214 I 242 F 238 M 238 M
280
P 215 P 243 P 239 P 239 P x
281
I 216 L 244 I 240 I 240 I
282
N 217 N 245 N 241 N 241 N x
283
E 218 E 246 E 242 E 242 E x
284
P 219 E 247 P 243 P 243 P
285
V 220 S 248 A 244 A 244 A
286
H - 249 L 245 P 245 P
287
G - 250 A 246 G 246 G
288
T 221 S 251 K 247 K 247 R
289
K 222 K 252 K 248 K 248 K x
290
R 223 G - -
291
K 224 A 253 K 249 K 249 K
292
S 225 G 254 S 250 S 250 S
293
Q 226 Q 255 Q 251 Q 251 Q x
294
I 227 I 256 I 252 I 252 I x
295
Q 228 E 257 D 253 Q 253 Q
296
T 229 E 258 E 254 E 254 E
297
Y 230 F 259 Y 255 Y 255 Y
298
L 231 L 260 L 256 V 256 V
299
E 232 M 261 E 257 D 257 D
300
H 233 Q 262 F 258 Y 258 Y
301
N 234 F 263 Y 259 N 259 N
302
E 235 N 264 G 260 G 260 G
303
G 236 G 265 G 261 G 261 G x
304
A 237 E 266 A 262 A 262 A
305
G 238 G 267 G 263 G 263 G x
306
L 239 I 268 V 264 V 264 V
307
Q 240 Q 269 Q 265 Q 265 Q x
308
H 241 H 270 H 266 H 266 H x
309
L 242 V 271 I 267 I 267 I
310
A 243 A 272 A 268 A 268 A x
311
L 244 F 273 L 269 L 269 L
312
M 245 L 274 N 270 K 270 R
313
S 246 T 275 T 271 T 271 T
314
E 247 D 276 G 272 E 272 E
315
D 248 D 277 D 273 D 273 D x
316
I 249 L 278 I 274 I 274 I
16
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imagen33
imagen34
imagen35
imagen36
imagen37
imagen38
imagen39
imagen40
imagen41
imagen42
imagen43
imagen44

Claims (1)

  1. imagen1
    imagen2
    imagen3
ES13164699.4T 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso Active ES2594284T3 (es)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP10190629.5A EP2453012B1 (en) 2010-11-10 2010-11-10 HPPD variants and methods of use
EP13164699.4A EP2669372B1 (en) 2010-11-10 2010-11-10 Hppd variants and methods of use

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ES2594284T3 true ES2594284T3 (es) 2016-12-19

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ES13164699.4T Active ES2594284T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso
ES13164702.6T Active ES2588991T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso
ES10190629.5T Active ES2588918T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso
ES13164696.0T Active ES2588802T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso

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ES10190629.5T Active ES2588918T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso
ES13164696.0T Active ES2588802T3 (es) 2010-11-10 2010-11-10 Variantes de HPPD y procedimientos de uso

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CN110616203B (zh) 2018-06-04 2022-04-01 青岛清原化合物有限公司 突变型对羟苯基丙酮酸双氧化酶、其编码核酸以及应用
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CN114364793B (zh) * 2021-01-15 2024-07-23 江苏省农业科学院 具有除草剂抗性的hppd突变型蛋白及其应用
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