CN1661079A - Correlation between catalase gene and essential hypertension - Google Patents

Abstract

A process for testing the susceptibility of primary hypertension includes detecting if there are variations in hydrogen peroxidase gene CAT, transcript and/or protein of an individual, and determining its high susceptibility if there are. Its reagent kit is also disclosed.

Description

Correlation between catalase gene and essential hypertension

technical field

The present invention relates to the fields of molecular biology and medicine. More specifically, it involves the single nucleotide polymorphism (single nucleotide polymorphism, SNP) of the catalase gene (catalase, CAT) and its correlation with essential hypertension. The present invention also relates to methods and kits for detecting these SNPs.

Background technique

Hypertension refers to a group of clinical syndromes with elevated systolic or diastolic blood pressure. There is an obvious causal relationship between the increase of blood pressure and the occurrence of coronary heart disease, renal dysfunction, hypertensive heart disease and hypertension complicated with stroke. The latest diagnostic criteria for hypertension are systolic blood pressure ≥ 19kpa (140mmHg) or diastolic blood pressure ≥ 12kpa (90mmHg), and those who meet one of them can be diagnosed as hypertension.

Most hypertensive patients only have mild subjective symptoms in the early stage of high blood pressure, such as headache, dizziness, insomnia, tinnitus, irritability, difficulty in concentrating work and study, and fatigue. With the development of the disease, especially when complications occur, the symptoms gradually increase and become more obvious, such as numbness and stiffness of fingers, pain in the lower limbs when walking more, or muscle pain and tension in the neck and back. When there is palpitation, shortness of breath, chest tightness, and precordial pain, it indicates that the heart has been involved. When there is nocturnal frequent urination, polyuria, and light urine, it indicates that the kidneys are involved and the renal arterioles are hardened. If a hypertensive patient suddenly develops confusion, deep and irregular breathing, and incontinence, etc., it may indicate cerebral hemorrhage. If one side of the body gradually becomes inconvenient, numb or even paralyzed, it should be suspected whether there is a cerebral thrombosis.

There were no obvious abnormal signs in the early stage of hypertension. Abnormal signs may appear when the brain, heart, kidney and other vital organs are slightly damaged. Common heart abnormalities include leftward shift of the apical beat, lifting-like pulsation sensation in the precordial area, enhanced first heart sound in the apical area, enhanced second heart sound in the aortic valve area, and systolic murmurs and diastolic murmurs. Arteriosclerosis and left ventricular hypertrophy, and a galloping rhythm in the apex may indicate heart failure. In addition, earlobe creases, capillary pulsation, hard or pulseless radial artery, and intermittent claudication of the lower extremities are also common.

In addition, due to certain predisposing factors or the development of hypertension itself, some hypertensive patients may have a significant or sudden increase in blood pressure, accompanied by damage to the brain, heart, kidney, retina and other important organs, which is seriously life-threatening, and a series of Special clinical signs are called hypertensive emergencies. The incidence of hypertensive emergencies accounts for 5% of the hypertensive population, common hypertensive encephalopathy, cerebral hemorrhage, acute left heart failure, clonidine acute withdrawal syndrome, acute myocardial infarction, rapidly progressive malignant hypertension, etc.

About 5% of hypertensive patients have no symptoms, do not know when blood pressure rises, and do not know when complications of blood vessel and organ damage have occurred. have high blood pressure. Therefore, finding out the genetic cause of hypertension, early prevention and detection can effectively control the incidence of hypertension and minimize the damage of the disease to the human body.

Essential hypertension (essential hypertension, EH), also called hypertension, is an independent disease with its own etiology, occurrence, development, outcome, and clinical manifestations. Clinically, it is mainly manifested as an increase in arterial blood pressure. Accounting for more than 90% of hypertensive patients in the population, the pathogenesis is not fully understood at present, and it is mainly diagnosed as essential hypertension (hypertension) only after the hypertension caused by other diseases has been ruled out. The increase in arterial blood pressure is mainly due to the increase in the resistance of peripheral small arteries, and at the same time there are varying degrees of increase in blood volume and cardiac output. In the late stage, the heart, brain, kidney and other organs are often involved, and serious complications such as hypertension, heart disease, heart failure, renal dysfunction, and cerebral hemorrhage occur. The treatment of essential hypertension is mainly to lower blood pressure while preventing the occurrence of complications. The causes of death in patients with essential hypertension are cerebrovascular accident, cardiovascular accident and renal insufficiency. In my country, cerebrovascular accident is more common, followed by heart failure and uremia. In European and American countries, heart failure is more common, and cerebrovascular accident is more common. and uremia followed.

As the most common cardiovascular disease, the incidence of essential hypertension is increasing year by year, and it can cause serious heart, brain, and kidney complications. It is a major risk factor for stroke and coronary heart disease. EH is a polygenic disease caused by the interaction of genetic factors and environmental factors. Finding EH-related genes and clarifying the genetic mechanism of hypertension has become a research hotspot.

Although there have been many studies on various gene polymorphisms and essential hypertension, there is no report confirming the correlation between the CAT gene and essential hypertension, and there is no confirmation that the SNP of the CAT gene is associated with essential hypertension sex reports.

To sum up, in order to finally realize the treatment of hypertension, there is an urgent need in this field to find essential hypertension susceptibility genes, and to develop methods, kits, and related therapeutic drugs for detecting essential hypertension.

Contents of the invention

The object of the present invention is to provide a method and detection kit for diagnosing (especially early diagnosis) hypertension.

Another object of the present invention is to provide a new method for treating hypertension.

Another object of the present invention is to provide a pharmaceutical composition for treating hypertension.

In a first aspect of the present invention, a method of diagnosing susceptibility to hypertension in an individual is provided, comprising the steps of:

detecting the individual's CAT gene, transcript and/or protein, and comparing it with normal CAT gene, transcript and/or protein,

A difference indicates that the individual is more likely to have high blood pressure than the normal population.

In another preferred example, the method detects the gene or transcript of CAT, and compares the difference with the normal CAT nucleotide sequence.

In another preferred example, the difference is the following single nucleotide polymorphism:

26328-bit T→C;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In a second aspect of the present invention, a method for detecting whether there is a single nucleotide polymorphism of catalase gene CAT in a sample is provided, comprising steps:

(a) amplifying the CAT gene of the sample with CAT gene-specific primers to obtain an amplified product; and

(b) Detect whether the following single nucleotide polymorphisms exist in the amplification product:

26328-bit T→C;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In another preferred example, the gene-specific primers have the sequences of SEQ ID NO: 2 and 3.

In another preferred example, the length of the amplified product is 100-3000bp, and contains the 26328th position in SEQ ID NO:1.

In the third aspect of the present invention, a kit for detecting hypertension is provided, which includes primers for specifically amplifying the CAT gene or transcript, more preferably, the amplified length of the primers is 100-3000bp, And contain the amplified product at position 26328 in SEQ ID NO:1.

In another preferred embodiment, the kit also contains reagents selected from the following group:

(a) a probe that binds to the mutation at position 26328 in SEQ ID NO: 1;

(b) Recognizing the mutant restriction enzyme at position 26328 in 8EQ ID NO:1.

In another preferred example, the mutation is selected from the following single nucleotide polymorphisms:

26328-bit T→C;

Wherein, the nucleotide position numbers are based on SEQ ID NO:1.

In the fourth aspect of the present invention, a pharmaceutical composition is provided, which contains a safe and effective amount of CAT protein and a pharmaceutically acceptable carrier.

Detailed ways

After intensive and extensive research, the inventors have determined and analyzed the SNPs of a large number of candidate genes. For the first time, it was discovered and proved that some SNPs of the CAT gene are closely related to hypertension, and its new function was discovered: the change of CAT will lead to hypertension. C) There is a significant difference in the distribution between the case and the control group (P<0.05), so it can be used as a specific SNP for detecting hypertension. The present invention has been accomplished on this basis.

CAT gene

The detailed sequence of catalase gene (catalase, CAT) can refer to the nucleotide sequence that accession number is AL035079 (can refer to website http://www.ncbi.nlm.nih.gov/), as SEQ ID NO: 1 shown.

The catalase (CAT) gene belongs to a peroxidase whose structure and function have been clarified. In 1986, Quan et al. isolated and characterized this gene in humans (Quan et al. Nucleic Acids Res. 14:5321-5335, 1986.). The full length of the gene is 34Kb, consisting of 13 exons. The gene product is composed of 60Kd four subunits, each subunit contains a heme group, and the molecular weight is 240Kd. Catalase can decompose toxic H ₂ O ₂ into H ₂ O and O ₂ to protect cells from oxidative stress. The protein was first isolated from red blood cells in mice and subsequently in red blood cells in humans. Catalase, SOD and glutathione reductase constitute the main antioxidant system in cells.

Catalase is a ubiquitous protease present in currently known organisms, and is expressed in high abundance in liver, kidney cells and erythrocytes. Catalase protein, SOD and glutathione reductase together constitute the intracellular antioxidant system, and catalase plays a major antioxidant role in red blood cells. Studies have shown that Japanese catalase deficiency is related to the low concentration of catalase in red blood cells, and the concentration of catalase in red blood cells of this disease is only 0.2-4% of normal people (Ogata, M. Aatalasemia. Hum. Genet. 86:331-340; 1991), and is due to a splicing mutation (fifth base G→A of the fourth intron). In the Hungarians, polymorphisms were found in the upstream regulatory region (-21, A→T, the translation start site is 1; (Goth.L et al.Electrophoresis.1997jun:18(7):1105-8). So far, there are few common polymorphisms recognized by everyone about the catalase gene, and most of them are polymorphisms with ethnic characteristics.

Years of research have shown that oxidative stress imbalance can lead to diseases such as atherosclerosis and some aging diseases (Jose M et al., Biochemistry, 32(8), 595-603, 1999). As the main antioxidative enzyme in erythrocytes, catalase plays an important role in antioxidation (Gaetani GF, et al., Blood 87: 1595, 1996).

In 2000, Goth L and Eaton JW found that catalase deficiency would lead to a higher incidence of diabetes (Goth et al, Lancet 2000 25; 356(9244): 1820-1).

Since catalase can degrade toxic H ₂ O ₂ to avoid the harm of oxidative stress to cells, it was screened as a candidate gene for hypertension in an isolated population in China.

The inventors sequenced almost the entire region of the CAT gene and found many SNPs, most of which were not associated with susceptibility to hypertension, but association studies showed that position 26328 T→C was associated with susceptibility to hypertension Very high SNP. The SNP is located in the first intron of CAT, suggesting that it affects the post-transcriptional splicing process of CAT, which in turn affects the expression of catalase, and ultimately leads to a significantly higher susceptibility to hypertension in carriers than in normal populations.

Based on the new discovery of the present invention, the CAT protein or polypeptide has many new uses. These uses include (but are not limited to): direct use as drugs to treat diseases caused by CAT protein function hypofunction or loss (such as hypertension), and for screening substances that promote the function of CAT protein, such as antibodies, polypeptides or other ligands. Screening the polypeptide library with the expressed recombinant human CAT protein can be used to find therapeutically valuable polypeptide molecules that can stimulate the function of the human CAT protein.

On the other hand, the present invention also includes polyclonal antibodies and monoclonal antibodies, especially monoclonal antibodies, specific to human CAT DNA or polypeptides encoded by fragments thereof. Here, "specificity" means that the antibody can bind to human CAT gene product or fragment. Preferably, it refers to those antibodies that can bind to human CAT gene products or fragments but do not recognize and bind to other irrelevant antigen molecules. Antibodies in the present invention include those molecules capable of binding and inhibiting human CAT protein, as well as those antibodies that do not affect the function of human CAT protein.

The present invention includes not only complete monoclonal or polyclonal antibodies, but also immunologically active antibody fragments, such as Fab' or (Fab) ₂ fragments; antibody heavy chains; antibody light chains; genetically engineered single-chain Fv molecules; or chimeric antibodies.

Antibodies of the present invention can be prepared by various techniques known to those skilled in the art. For example, purified human CAT gene products, or antigenic fragments thereof, can be administered to animals to induce polyclonal antibody production. Similarly, cells expressing human CAT protein or antigenic fragments thereof can be used to immunize animals to produce antibodies. Various adjuvants can be used to enhance the immune response, including but not limited to Freund's adjuvant and the like.

Antibodies of the invention may also be monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology. The antibodies of the present invention include antibodies capable of blocking the function of human CAT protein and antibodies that do not affect the function of human CAT protein. Various antibodies of the present invention can be obtained by conventional immunization techniques using fragments or functional regions of human CAT gene products. These fragments or functional regions can be prepared using recombinant methods or synthesized using a polypeptide synthesizer. Antibodies that bind to unmodified forms of the human CAT gene product can be produced by immunizing animals with gene products produced in prokaryotic cells (e.g., E. coli); antibodies that bind to post-translationally modified forms (such as glycosylated or phosphorylated Proteins or polypeptides), which can be obtained by immunizing animals with gene products produced in eukaryotic cells (such as yeast or insect cells).

Antibodies against human CAT protein can be used in immunohistochemical techniques to detect the amount and/or mutation of human CAT protein in biopsy specimens. A preferred anti-CAT antibody is an antibody that does not recognize normal CAT but recognizes mutant CAT, or an antibody that recognizes normal CAT but not mutant CAT. Using these antibodies, the detection of hypertension susceptibility at the protein level can be conveniently performed.

Using the CAT protein of the present invention, substances that interact with the CAT protein, such as inhibitors, agonists or antagonists, can be screened out through various conventional screening methods.

When the CAT protein of the present invention and its antibody, inhibitor, agonist, antagonist, etc. are administered (administered) therapeutically, various effects can be provided. Generally, these materials can be formulated in a non-toxic, inert and pharmaceutically acceptable aqueous carrier medium, wherein the pH is usually about 5-8, preferably about 6-8, although the pH value can be changed according to the Depending on the nature of the substance formulated and the condition to be treated. The formulated pharmaceutical composition can be administered by conventional routes, including (but not limited to): intramuscular, intravenous, or subcutaneous administration.

Normal CAT polypeptides can be directly used in the treatment of diseases, for example, in the treatment of hypertension. When using the CAT protein of the present invention, other agents for treating hypertension can also be used simultaneously.

The present invention also provides a pharmaceutical composition, which contains a safe and effective amount of the CAT protein of the present invention and a pharmaceutically acceptable carrier or excipient. Such carriers include, but are not limited to: saline, buffer, dextrose, water, glycerol, ethanol, and combinations thereof. The pharmaceutical formulation should match the mode of administration. The pharmaceutical composition of the present invention can be prepared in the form of injection, for example, by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably manufactured under sterile conditions. The active ingredient is administered in a therapeutically effective amount, for example about 0.1 microgram/kg body weight to about 10 mg/kg body weight per day. In addition, the polypeptides of the invention can also be used with other therapeutic agents.

When using the pharmaceutical composition, the safe and effective amount of CAT protein or its antagonist, agonist is administered to the mammal, wherein the safe and effective amount is usually at least about 0.1 μg/kg body weight, and in most cases no more than about 10 mg/kg body weight, preferably the dosage is about 0.1 μg/kg body weight to about 100 μg/kg body weight. Of course, factors such as the route of administration and the health status of the patient should also be considered for the specific dosage, which are within the skill of skilled physicians.

Polynucleotides of human CAT protein can also be used for various therapeutic purposes. Gene therapy technology can be used to treat abnormalities in cell proliferation, development or metabolism due to non-expression of CAT protein or expression of abnormal/inactive CAT protein. The method for constructing a recombinant viral vector carrying the CAT gene can be found in existing literature (Sambrook, et al.). In addition, the recombinant human CAT gene can be packaged into liposomes and then transferred into cells.

The methods for introducing polynucleotides into tissues or cells include: directly injecting polynucleotides into tissues in the body; or first introducing polynucleotides into cells in vitro through vectors (such as viruses, phages, or plasmids, etc.) , and then transplant the cells into the body, etc.

The invention also relates to a diagnostic test method for quantitative and localized detection of human CAT protein level. These assays are well known in the art and include ELISA and the like.

A method for detecting whether there is a CAT protein in a sample is to use a CAT protein-specific antibody for detection, which includes: contacting the sample with a CAT protein-specific antibody; observing whether an antibody complex is formed, which means that the antibody complex is formed CAT protein is present in the sample.

The polynucleotide of CAT protein can be used for the diagnosis and treatment of diseases related to CAT protein. In terms of diagnosis, the polynucleotide of CAT protein can be used to detect the expression of CAT protein or the abnormal expression of CAT protein in a disease state. For example, the CAT DNA sequence can be used for hybridization of biopsy specimens to determine the abnormal expression of CAT protein. Hybridization techniques include Southern blotting, Northern blotting, in situ hybridization, and the like. These technical methods are all open and mature technologies, and relevant kits are available from commercial sources. Part or all of the polynucleotides of the present invention can be immobilized as probes on microarrays or DNA chips (also known as "gene chips") for analysis of differential expression of genes in tissues and gene diagnosis. RNA-polymerase chain reaction (RT-PCR) in vitro amplification with CAT protein-specific primers can also detect the transcripts of CAT protein.

Detection of mutations in the CAT gene can also be used to diagnose high blood pressure. Detection can be against cDNA or genomic DNA. The form of CAT protein mutation includes point mutation, translocation, deletion, recombination and any other abnormality compared with the normal wild-type CAT DNA sequence. Mutations can be detected using established techniques such as Southern blotting, DNA sequence analysis, PCR and in situ hybridization. In addition, mutations may affect protein expression, so Northern blotting and Western blotting can be used to indirectly determine whether a gene has a mutation.

The most convenient method for detecting the SNP of the present invention is to amplify the CAT gene of the sample with CAT gene-specific primers to obtain the amplified product; then detect whether the following single nucleotide polymorphism exists in the amplified product: 26328 T → C, wherein the nucleotide position numbering is based on SEQ ID NO:1.

It should be understood that after the present invention first reveals the correlation between the SNP of the CAT gene and hypertension, those skilled in the art can easily design an amplification product that can specifically amplify the position of the SNP, and then through methods such as sequencing Determine if there are 26328 bits T→C. Usually, the length of the primer is 15-50bp, preferably 20-30bp. Although it is preferred that the primer is completely complementary to the template sequence, those skilled in the art know that it can also be specifically amplified (that is, only amplify the desired fragment). Kits containing these primers and methods using these primers are within the scope of the present invention, as long as the amplified product amplified by the primers contains the corresponding position of the SNP of the present invention. A preferred primer pair has the sequences of SEQ ID NO: 2 and 3.

Although the length of the amplified product is not particularly limited, generally the length of the amplified product is 100-3000 bp, preferably 150-2000 bp, more preferably 200-1000 bp. These amplified products should all contain the 26328th position in SEQ ID NO:1.

Because the SNP of the present invention has a very high correlation with hypertension, it can not only be used for early and more accurate diagnosis of essential hypertension, but also can take precautions to make the carrier take reasonable preventive measures before the onset of the disease (such as in the Therefore, it has extremely important application value and social benefits.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific conditions in the following examples, usually according to conventional conditions such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's instructions suggested conditions.

Example 1

1.1 Research object

Since blood pressure is a quantitative trait that is affected by multiple genes and is affected by various environmental factors, it is necessary to consider the issue of genetic heterogeneity when analyzing related genes. Selecting isolated populations with a relatively single genetic background as materials for the study of polygenic diseases will help reduce the impact of genetic heterogeneity. reduce. Selecting such a group, whether it is used for linkage disequilibrium analysis or selecting families for family analysis, will help to improve the detection sensitivity of hypertension susceptibility loci. Therefore, in this embodiment, the isolated population is selected as the research object (the isolated population has the advantages of relative consistency of genetic background and relatively small number of founders).

On the basis of informed consent, 324 individuals of Han nationality over the age of 50 were randomly collected from Xiangchang Town, Dabie Mountain, Yuexi County, Anhui Province. 80% of individuals have only 6 surnames, the number of founders should be relatively small, and individuals of the same gender come from the same founder. The selected hypertensive samples require that the systolic blood pressure is not lower than 140mmHg, and the diastolic blood pressure is not lower than 90mmHg, and the blood pressure is measured twice to take the average value.

11% of them are located at the top of the blood pressure distribution (37 individuals; SBP>178mmHg) and 7% are located at the bottom of the blood pressure distribution (22 individuals; SBP<104mmHg), a total of 59 individuals were selected by direct sequencing method Perform SNP detection.

1.2 Experimental methods and results

1.2.1 DNA extraction

DNA was extracted from human blood by the conventional phenol-chloroform method, and the concentration was corrected to 20ng/ul for conventional PCR amplification.

1.2.2 Design of PCR and sequencing primers

According to the genome sequence of CAT in GenBank, the following primers were designed and synthesized. The specific primers are shown in Table 1 below.

Table 1 Primer sequence list Primer name Sequence (5'-3') SEQ ID NO: sense primer gcatgcacaactctctcagg 2 antisense primer ggtttctggggcatcactaa 3

1.2.3 PCR amplification of CAT gene

Using the extracted DNA as a template, PCR amplification was performed on a GeneAmp 9700 PCR instrument with the Touchdown program using Taq enzyme. The reaction conditions are: pre-denaturation at 94°C for 2 minutes, denaturation at 94°C for 30 seconds, annealing at 63°C for 40 seconds, extension at 72°C for 40 seconds, a total of 10 cycles, and the annealing temperature decreases by 0.5°C for each cycle; after denaturation at 94°C for 30 seconds, Anneal at 58°C for 40 seconds, extend at 72°C for 40 seconds, a total of 30 cycles; finally extend at 72°C for 7 minutes. PCR amplification products were verified by agarose gel electrophoresis.

As a result, an amplification product of 347 bp was obtained.

1.2.4 SNP discovery and detection

After the PCR products were purified by Resin resin, ABI-PRISM ^TM 377 DNA sequencer (appliedbiosystems (ABI)) was used for bidirectional sequencing with fluorescently labeled terminal termination method, and Polyphred software (University of Washington, USA http://droog. mbt.washington.edu/Polyphred.html) for sequence interpretation and SNP confirmation.

As a result, the following SNP was found to exist: 26328 position T→C.

1.2.5 SNP genotyping and association analysis

SNP genotyping by direct one-way sequencing. That is, typing and correlation analysis were carried out in hypertensive patients and normotensive control groups.

After the allele frequencies are counted, chi-square test is carried out. By comparing the individual data of the highest 11% with the lowest blood pressure of 7%, it is preliminarily determined whether the allele frequencies are linked with the blood pressure level.

It was found that the frequency of 26328-type C SNP in SEQ ID NO: 1 was 65.5% in hypertensive individuals, and 47.7% in hypotensive individuals. The frequency difference between the two is 17.8%, which confirms that there is a correlation between the 26328 C/T type SNP in SEQ ID NO: 1 and the occurrence of hypertension.

Example 2

Essential Hypertension Susceptibility Detection Kit

As described in Example 1, the mutation of T→C at position 26328 in SEQ ID NO: 1 is closely related to hypertension. Therefore, CAT gene-specific primers can be designed based on this mutation and detected by amplification using the patient's DNA as a template.

Prepare a test kit (100 person-times), which contains: name sequence concentration sense primer SEQ ID NO: 2 100pmol antisense primer SEQ ID NO: 3 100pmol PCR reaction solution Containing Taq Enzyme dNTP Magnesium Ion PCR Reaction Buffer

Take 3ml of blood from the male patient to be tested, and use a conventional method (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the high blood pressure detection kit to 1μmol/μl, and use the extracted DNA as a template to perform a PCR reaction with the provided primers. After the PCR products were purified, ABI-PRISM ^TM 377 DNA sequencer was used for bidirectional sequencing by fluorescence-labeled end termination method, and Polyphred software was used for sequence interpretation and SNP confirmation.

Alternatively, the chromatographic analysis of the amplified product and the normal control is performed with a denaturing high-performance liquid chromatograph (DHPLC), and the 26328 position T→C can also be detected.

As a result of the test, the susceptibility to hypertension of the subjects including 26328 T→C was higher than that of the normal population.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Sequence Listing

<110> Fudan University

  Shanghai Human Genome Research Center

<120>Relationship between catalase gene and essential hypertension

<130>040166

<160>3

<170>PatentIn version 3.1

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gatctcccct cctcagcctc ccaaagtgct gggcttacag gcatgaacca ccgtgcctgg 60

ccgaattaat gtgtttttaa accaaaatcc tgacatttat ttgaaacaaa ctaggcaatc 120

cttcacagtg cttctaaaag cccagaccaa gccctccttt tagaagagca agtctcatga 180

gcagcgtgag gtcagagatg atcaggtcca tgcacatttg tgtctttcca caatgtctga 240

ctttcttgaa gtaatctcaa tcacaaaagc cacgagctgc acggagttcc taaggaggca 300

gtttgcctca gtgcttccca tttgctcgat agtcagagct gcagtcacac aggctcaagc 360

cactccacaa gtcagtcaat gtggcagatc atacataata gtatacttaa aatataaatg 420

ttatagatta aacatgccac aacaaagtaa tattgaacat caaaagaaga aggataggaa 480

gaaagggtta atgaatcaat tcagggagag cgactaaggc aaaaagaatc tcctggtctg 540

cgccaggcag tccgagagcc ttcagtggca gatgccaggt gattatcatg attaacagca 600

agatggtgta gttaagacgg ccatttcaag ctgcgcaagt cctgttcttt ttatagccac 660

agtcctctag tgaggactga gagtggaaaa gtgtgcttgt tgatgtcctc atctggttgg 720

atgcagtctt tatttattaa gcaaacatct tgtccctgtt cgcaaagtgc cctatgaaat 780

ataagatgga gtctctaaga tggagttact tatgtcaagg ggcttgacac agcaaaagaa 840

cagattttta tatattaaga gtctaatatt tacattttta gatagtggtg tggcagagtg 900

taggtgaaaa acaccttttt ttttttttga gacggagtct cgctctgtca cccaggctgg 960

agtgcagtgg cgcaatcggc tcactgcaac ctccgcctcc caggttcaag caattttcct 1020

gcctcagcct cctgactggc tgggattact ggcgcctgcc accacgccca gctaattttt 1080

ttttgtattt ttattatagaga tggggtttca ccatagtggc caggctgctc ttgaactctt 1140

gacctcgtga tccgcccgcc tcagccttcc aaagtgctgg cattacaggt ggcagccacc 1200

gcacccagcc agaaaaacac cttttaagat gagaatgcaa gaaatgaaaa tgttatgtcc 1260

acacaaaaat atgtacagaa atgttcatac cagcactatt cataatagcc aaaaagtaga 1320

aacaatccaa atgtccatca aatgataaac ggataaataa aacatagtaa atccataaaa 1380

tggaatatta tttggcaatt aaaaaaatga agcactcata gatgctacaa tatgtaagaa 1440

tcttgaaaat attatgctaa gtgaaagaag ccagtcacaa aagaccacat actatattat 1500

tccattaaa tgaaacatgc agagtaggca aatcttagat tcatgatggc ctagggctgg 1560

ggactaggtt aagaaagaaa tggagagtgg ctgctgatag gtacaggttt ccctttgggg 1620

tgatgaaaat gctccaaaat tgattgtggt gatggttgcg caattctgtg aatatacaaa 1680

aaaaatttac tcgtacactt taaatgggtg gtttttacgg tatgtgaatt atatctcaat 1740

aaagctctta ttttaaaaaa gagagagaag aaaaaaggaa gaaggaagaa aaaaaaagga 1800

ggaggatgag ggcaaggaca agaagaagag aggagggag gaggaagaaa aatgaggagg 1860

aggagaaagg gacgaggagg aaagctgggc atggtgatgt gtgcctgtag tcccagttac 1920

tcaggaggca gaggccaccc tgggcaacat aatgatatgc cgtctctaaa ataaataaac 1980

aatgaagaag gcagtgtgaa cgaccaactg tgtcaaacac cgctgagagg tcatgtaaga 2040

caaggactaa gaatggacca ttggattttt ttgtggggta aaatatacac aaaacttacc 2100

atttagccat ttttaggtgt acagttcagt agcattaaat acattcacac tgttgtgcaa 2160

ccatcaccac caaccatctc cagaagttct cacctttccc aataaaaagt gcagtggcgc 2220

aatcttggct cactgcaacc tccacctcct gggttcaagc gattctccca cctcagcctc 2280

acgagtagct gggattacac gcacatgcca ccacacccag ctaatttttg tatttttagt 2340

agaaatgggg tttcaccacg ttggccaggc tgatctcaaa ctcctgacct caggtgttcc 2400

acccgccttg gcctcccaaa gtgctgggtt tacaggcgtg agccactgca cccagcccga 2460

tatatctatt ttctatgtct acaaatataa cagtctcaaa gcatactcaa atacgggggg 2520

aattaggcag gagggtttga cagaggagta aagaaatttg agctaaaaga ccaagaatga 2580

gtttgtatct gtctgctagt ggtctgtttc ctgaagcccg taactcagga actcctgaaa 2640

gtataaactg gtttggtttt ttgtttttag ttgttattgt tgttgttttc agacaaggtc 2700

tcactctgtt gcccagactg gagtgcagtg gtgcaatcac gactcactgc agcctcaacc 2760

tcctgggctc aagagatcct cccgcctcag cctccctagt aactgggact acaggcatgc 2820

gttaccatac tcggctaatt tttgtatttt ttgtacagac agggcttcgc catgttgccc 2880

cggctggtct caaactcctg agttcaagtg atcctcctgc cttggcttcc caaagtgctg 2940

ggtttacagg catatgagcc accacacccg gcacaaactg gttttgaata tgaacggaat 3000

aaagattttc aaggaaaaca aattttcatt tagtattcag aactcatctt taaagtcttc 3060

catgtgattt tcaaccatga taggcaaata tgggctttaa aaagaaggaa gaggaaagtg 3120

accagtggtt ctgagccctt gttttaaaaa gtcaatcaca ggtaagtgag caaatggaca 3180

agagatttaa gtaattttca aagttggggc atttgctatt ttatcagtgt ctcagaaggc 3240

tgtccaaagg ctatagattt aaacaaataa catggagtcc tcgagatact ggcatttcca 3300

tttttaaggg taaacatctg aaaatgatga tgttcaaatt aagagtctgg tagcagttta 3360

cagttacata tggagtcgaa agaaagaaaa taccatctct gttcttacgt acagattat 3420

tattgtaaaa atgtaagcaa ttacaaattg atctgttgtg aaatcagtgc tgaattaaca 3480

ttaatgtaac tccagtgaaa aaaatgaatt tgatcaatta ttatcaggaa taaacattaa 3540

gccatgacgg tgctcaagtt atcttctgat gacactgctg tattaaatgt acttttccta 3600

tgagaacctt agaaaaataa aaatttctga gtagatcatc tttatctttt ctttttatat 3660

atttatagcc atattttaat ataataggcc atgaggtgat atgtaaacat acataatttc 3720

aacaaatttt gtcaaataaa atcatatttc tagctagaaa cttgccttct aatcatttta 3780

aatgaataat ccaatcatcc gtcaaaacaa attttttttt taaatgactg ttaaatccta 3840

accttcattt tcccctgggg aaatagcatt aaaattgtta cacaatctgc tatagaaagt 3900

gggattctct attttgaagc cactagcttg catttgcaca tctagcacag gagaatcttc 3960

atccagtgat gagcgggtta cacggatgaa cgctaagctt cgctgcacag gtgcagggcc 4020

ccggcctcac agatttgcct tctcccttgc cgccaagtga gatccggact gcacaaaggt 4080

gtgaatcgca ttctgtttta aaagaacaag atgagcaggt taatcacctc atgggcagca 4140

acgtgatatt tactcagttt taatatgtat gcaaatgaaa ataataattc agaattcatt 4200

cactttaact gtcttaaatt gcccagcgct aaaatgaaga gtatgtgttt tgccagtgaa 4260

ttctttatct ctcaaatgaa tttctgcaca gctgcctaag aaatctaaca gtggcaggta 4320

atggcccctg cagactgtaa tgctaagtgg taactaagtc atgcctactc caacccagca 4380

cacacagcct ctgcatggcc aggctgctcc cagcttacct taggcttctc agcattgtac 4440

ttgtccagaa gagcctggat gtggctcccg tagtcagggt ggacctcagt gaagttcttg 4500

acctaaggcc aaaggaaaat aaatgcaagc attackctaa ctccctacta tatcactccc 4560

cagggactgt ggtttcccag tgtttattcc ataggggagg aaagacgttt cagcaggagg 4620

cagtactaca ggtgtttaac aatgccagcc tcagagttaa gatgcaggtc cagtgaacac 4680

tctgacatat actggctgag caacctcgga taggtcactt aacttactga gcttatgtcc 4740

ctctgccgaa atgcgcaaga aaatgacacc taatcagtat aattttaaaa atactatttg 4800

gtctttgtcc cccattcctg acagatct aaaacccttc gaatgtactg tcttttatat 4860

actactaaga tgattcatgg gggcctacat agattcctga tggaggctca caccaggaag 4920

aacaagcacg cgatcagaag gttagaactt ctagccccac ccccaaacct ccagggatgg 4980

gagaggggct ggagattgta ttcaatcacc catagccaat aatttcatca atcatgccta 5040

tgtcataaaa cctccagaaa aacccacaac tggcaaggtc agggagtgtc tggattagtg 5100

cacacatcca ggagctagaa gggtggtcca ctcagggagg cacagaagct ccgaggcctt 5160

cccccatgcc cctcttccat ttgtctattc ctgagttgta ttctttataa taaattggta 5220

atcataagta aatcattttc ctgagttcta tgagtctttc cagcaaatta tcaaacctga 5280

ggaaagggtt tgatttgcag tcagaatctg cagaagtgtg catagccagg gtacctcatt 5340

tgttatctaa agtgggaaat gacccctaaa tctttgggtt ctgactctag ctccaggagc 5400

tactgttaga attgaattga gttataggac acacagttac tgtcaaagaa ttacagaatt 5460

gcttattgtc ggggaaaaaa acccatacaa cctaggccag gcgcagtggc tcacgcccga 5520

aatcccagca ctttgggagg ccaagatggg tggatcacct gaggtcagta gttcaagacc 5580

agcctggtta acatggcaaa acccccatctc tactaaaaac acaaaaatta gtcgggcatg 5640

gtggggcatg cctgtagtcc aagctactgg ggaggctgag gcaggaaaat ctcttgaacc 5700

caggaggcag aggttgcagt gagccgagat catgccactg cagtccagcc tgggtgacag 5760

aacaagactc catcttaaaa aaaaaaaacc cacacaacct acctcacagg atgctgtgag 5820

aaaaatgctg agaagtgagg tgcatttact gcatgaaaag agctgacctc agagtaagcg 5880

cttaacaaat tacagtgatt tacattgaga aacgggccca gacacagcct aaagttaatg 5940

tgaaggatac aagaacactg gtacatgcag cacaactcca cacacttcag gcaggaattc 6000

cccagtggac agcaagagga ggcagggcat tgggaaccca taaaagcctt acagtttcag 6060

catatgtgaa aggagaatgc caagggttcc agcccagttc tgctatttac aagctgtcgt 6120

atccattctc tccagtaaaa ggagataaat cctgccctgc ctaccacaca gagcgcctgt 6180

caagctctct gctgatactg gaacccaggc tctgggacac agcccagaca gctgctagcc 6240

cagctgcccc taggaactga gaagcagcaa ctaggggcca aggaaaaagg gggatttcta 6300

gatttgcaaa aagcttagct ctgggacagg ccaggaaacc agaagtgaga gggcttcact 6360

cagctcgatg agcggccatc agaaataccc cagcacctta gcatgaaagt ctagcctgcc 6420

ttttggaagc attgcagttc ttattccaga aatcatgcaa atttctagtc tgcatctgaa 6480

ggaggatcga ggtattatga cattaagcct ttaggcaggt tttaaaccat aaaaatgcaa 6540

aaaagaaaga cagactgatt gactgattga ctgattttag agacagggtc tccctatgtt 6600

gtccaggctg gccctgtatt cctgggctca agcgatcctc ccatctcaac ctcctaagta 6660

gctaggacta caggtgcatg ccatcacacc ggctgatata tttattatca tccaaagttt 6720

aagtagaatt gcagcaggac tcacattttg atattgcaca tttactatct tgcacagaac 6780

ttggctgcag ctgaattaaa cagaatagga caaaattcag tctgaacaca actaaattag 6840

tacacactgg acaaacagct aaggacgatg gatatgccag accagataaa acatggaaag 6900

aaaataatca gaagttggtt ggggattact agtttatttt ttttttttta agttctctta 6960

agtcaagtaa aacagaaatg gcactttctt tttagggttc tcccacacag ctgcactgtc 7020

ttccttagcc agcagaggac acccttcagc ttacaaagac tcaccgcttt cttctggatg 7080

aaaatttgtg catccttcag gtggccggca atgttctcac acagacgttt cctctgttcc 7140

tcattcagca cgttcacata gaatgcccgc acctgctcag agaaaagagc aagtgcagag 7200

aattgaatca ccagtttatc accaacaaaa ttcacctact acaacactta ggaaatcaat 7260

gataaaaaaa actttagaag ttaagaataa ttttaaattt taaatttggg ttgtgtcctt 7320

cactttttgt tttaagcaaa ataagtaaaa ttcatttgtt aatagctcat acctgctttc 7380

taataatatc ctttatagta actgctataa atctcttata aatagatcta caatttaaaa 7440

cctcaccaca ttcttaatac tgacaagtct aaaatgtgca agctgccttt gaatgtaaac 7500

ttagcacaat acctttgttc aattctccca gaaccatgga ggaaaattaa ctaatttagc 7560

tagagatata atttgatata tagataatta tatctatata tagacatcta aaaaactacc 7620

tcatttatac atgtgtatct gtagttatc agaagccaat atttcttaag caagacatat 7680

taatagtgat attttgctag attattgggt tgacctctaa aataaatatc cataaattaa 7740

aatgaaaaat aggagcttag tatcagatat gtgaatccga gcagcctctg ccccattctg 7800

aaatcaaaag gttgtagcta tgattttat cttacatttc attcattcat tccactcatt 7860

caacaaatat tgagtatcta taatgtgcga gactctgttc tacgtgctgt tctattaatg 7920

tttatgaagt ccaataatat ccaattatcc aaagtatcaa cttcataaaa tattaagaga 7980

tgcatcattt ccataagtca tcaaatatca ggctataatt atgtttttaa atttttgagt 8040

aaagtttaaa cctgatacca aagttaatca gacattttcg attaacacaa tctaatcaaa 8100

gtactatatc ctggggattt cctcagacct atctccaatt cactaattct ctcttcagct 8160

gtatttgaca tattgcgtga aaacatcgac tgggtcacta tatctgttga ctttttattt 8220

cacagattgt aatttttgtt tctagaaaat attatttggt tatttttcaa atctacctgt 8280

ttttctcctc atatcctatt gtcttaataa tattcctttt ttgcatctct ttaaatatgt 8340

atgtttattt taaagcctct ttccaattat catgtctatg attcttccat ctattattgc 8400

atgtgtaatt tctcattgga tttccttgta tgtattctaa tttttcacta tgactcatca 8460

gatgaagtag tcttgataac atctattgtt ctatcttcca attcactaat cttctcttca 8520

gccatgttaa cctcctttaa aagccatcta ttaaattcta atttcagttg ttacattttc 8580

tagttatgtc cattcaactc tttttctata gcttaaagtt ctcttactaa attcttcaag 8640

cagtcatcta ttttcttttt tttttttttttttttttttg agacggagtc tcgctctgtc 8700

gcccaggctg gagtgcagtg gcgcgatctc ggctcactgc aagctccgcc tcccgggttc 8760

acgccattct cctgcctcag cctcctgagt agctgggact acaggcgccc gccaccacgc 8820

ccggctaatt ttttttttgt atttttagta gagacggggt ttcaccgtgt tagccaggat 8880

ggtcttgatc tcctgacctc gtgatccgcc cacctcagcc tcccaaagtg ctggattac 8940

aggcgtgagc caccgcgccc agcccatcta ttttctcaaa cttattaatc agtcatttta 9000

aaacctgtat ctgacagttc catttattaa gcctccatta ggtttgtttc tactgtcagt 9060

ttgtaggggg ttttttctcc ctcttgattt tcagtcataa ggtcttaatg tgttaggtgc 9120

ctggtcattt attgttgaat gctgaacatt tccatataaa aaggtctaga atcatttgag 9180

tttctatttt ccaccaaaga aaattttgtt tgcttctgac agatggcgta gaaagagatc 9240

accttaatcc aatctgagac tgagcagagg caaagttatg tttcctgtat tgtgaaagca 9300

agtttatttc atgtttgccc ttattcctag gcatggccct ttaaggattt caactgaaag 9360

ccttgagtat aactagaggc tctgcttctt agcagacctt gaattccaat ttttgtccct 9420

tcagactcat gagactgctg taaacaccac tcagcttctg gaccagtgct tacaaattaa 9480

caaatacctt aagaggaaag gaaaccaaat gtcaagctta cttcactgca cttttttttc 9540

ctgcacttct tttccatctt tgatcttggt cccttaaatc cttgttgctt tgttagctct 9600

ccaaagcctt caaacaagtg ctttatgtat tttacctacc ttttcaatgt tctccatgaa 9660

acggtaagta agatacaagc tagtctgcca tagacagatg caagcccaag tgaatcattt 9720

ttaagagaat aaatctggac cttcttatgt atttcccatg tgatacatca tcacagaatt 9780

atgctatgaa tctataacaa tagtattgtg attacttaaa aaccacacta tatcctatat 9840

agtgataaga aattctctca ataagaatga gcacataata aattttcatg tgtaagcaat 9900

agcctcatta atttttgcag gggttgtatg agctggttta atacctgtat tgacctcaca 9960

taggcaaaat tctttgagag gcagggaatg tactgtaata tacttggtgt catctgcatc 10020

accaaaaata atgcagttac ataaaaggta tgcaggaaat gttttaagct catctaaaaa 10080

tggtttat cccacaataa tgtaatttct tttttatata tacgtatata tgtatatacg 10140

taaatgcttt tcaagcaaag atatcctcac agcgaattaa ttcatgtata agtatttttt 10200

acttggaacc cacattcaca aactgagaaa ctagaaatag gttctaatct aatttaaata 10260

ctaagtttaa cactaactgc tcaaacagag ctctagaaca actatacctt taactttat 10320

atttattgt gtgtggtata agttgcactg gacacattaa ggaggcagct gcccacctac 10380

agcccttcca agtccaaggg cctccttcat agcatgtgct gaaacagctc ttagggcagc 10440

cataggcaac tggaaagagg atgagcattt gccctggggc ggttaactag agcctgacca 10500

aggccaatgg tgcaacctgg ggtgaaaaga ttagccaagt caataggatt ccatctctct 10560

agaatctaga gatagacatg ttgagagaag gagtaaaagg tggtccaaaa aaaaaaagta 10620

ataatggaga aagacgccat aaaggagagt tttccattgt tgtggaagag ccacaaccat 10680

gataaaacag aagctctgac gcagatagga gaaaacagga agctgtggta aaaataatca 10740

attagcattg gcaagcagaa gcagacagat ggagaataag ccacgtgaag gacgagccct 10800

gcagtggaaa cgcccacact cttgtggccg aggctcctgg agctattttg gatctctagt 10860

tctagtcctt gaacctggac tgtcctgccc agattcttag atttgtctcc acgtaagctg 10920

tacatgctca tttcttaaat tcccatgaga taactgtttc cccaatgccc cctccccata 10980

tcccttcaat aaagtttgaa gaaactgaga cggagtctgc atgctttgag gcaaaagacc 11040

agagtaatat aacccttcaa ctccaccag taaggggccc aatttactat tacagatgag 11100

tcttccttga attcaattta agatacatca gacagttggg gcatttgcag ggagggggat 11160

tatgcaggca ttgacaagga gaattagcag gtgacttcca tagcagataa agagaagtca 11220

ttacctgagt aacgttatca tcattggcag tgttgaatct ccgcacttct ccagaatatt 11280

ggatgctgtg ctccagggca gaaggctgtt gttccggagc accaaagctg ttggggtagt 11340

aatttggagc accaccttca aaacaaaacc accgatgc acttaggaat aagaaaagtc 11400

aagtaagtct gcaaataatc actgtgatga gaaataaatt caagtggtaa tcaaatacac 11460

acgtacacat acagatat aaacacaacg cacacatatt atttttaatt tcccgcattt 11520

agctacaggt agtatgaaga ttcttaggga acgctgccat ctgctaaatg cttgcacagg 11580

ctgccgcact ttatgaatga atctcaaagt cccttttgac acttcaaagg gtctcatcta 11640

tcaagtcttc tgaggggtac agcacatgta gaagtttata caggcttgat ttggccacca 11700

tatagggatg aggctcaaag acaatttca aaattgactt gaaaaattgt gtcatttgga 11760

cttgggctca attcaaacac tgcatcacaa aattcttgag ggaaatctga aagttctggt 11820

tctgctcctt gaatctacag acacaagcat ggagctctgt gatgggacct gactgctgca 11880

aggtcacacg gtagccagtg gtagatatgg ggttggaact ctgggttttt tgtctcctcc 11940

agcccagcat atacagtatt tccattacag aactctactt tgctattccg taaagtgttt 12000

ttaggattct cagggtatct ctcctttact gacaatgatg actttgttcc attgtagctt 12060

cactttcata ccctccacag accaataaac aaagaatttt ttaaaatgtg gtaattgtat 12120

atcagcactt acttatctgg acttataaga aacagtgaca aatacaatca aaatattttc 12180

tatgactttt gaaataaatt ataaggcttt tttttcgttt tagcatattt attttctcaa 12240

ctatgctttg gctgacatta gcagaagctt tcattacccg agcacacact gcctaaacag 12300

gagtggccca gaaacatatt aagtggtttg agcaaaccaa tctggggttg gaattttagc 12360

cagctcactg ccgtaataga ttatccacag tggataatca agaggtgcct atgtatgaca 12420

ttcatcaaga caccaagtgt gtgtgtaggg gaaggaagaa caccataacta ttatagagcc 12480

caattattca ctcaacaaat tcaacagatc agtcacatct acatgaaaag caaaatgctc 12540

caaaaaccag ccatcacagt gttttcttca ttaagaacta gctctttgaa atgtgcctag 12600

ttaattttta aaatcctact tccaaatgag ccctgacctt tactcaaagt ggtagttgtc 12660

tttaaagcac tgaccacagg aacaattaag ctctttcact gagcttcagg gtcacagccc 12720

aaatgctcta cttttacaat caagccaggc gcggtggctc atgcctgtaa tctcagcact 12780

ttggggggcc aaagtgggtg ggtcacctga ggtcaagagt tcaggaccag cctggccaat 12840

gaaaccctgt ctctactaaa aatacaaaaa ttagctggcg tggtggcaca ctcctatact 12900

tccagctact cgggaggctg aggcatgaga atctcttgaa cccagaggca gaggctgcag 12960

tgagccaaga tcatgcactg cactccagcc tgggcaacag agtgagactc tgtctaaaaa 13020

ataaataaat agataaataa atatagcaaa atgtaatagc tatgaccttt gggtgttggg 13080

actatatggg aatttttctt ttttcttctt tttcctatat attccaaatt ttctttagga 13140

cacttgtatt atttttatca tagaaaaata aactttgttt gaatatagac atctgtggct 13200

ataaatataa atcatcagta aaatttctaa agggatcaat gcatgcagtc tatattacta 13260

aaacacgtgt cctaaatggg aaaagaaaat aactaagatc tatattaaac tgtggaatag 13320

taaaaggaaa aaactttaat caagcattct gcttgactta ttagaaaaag taattagaac 13380

cagaggctgt ttgtatacga caccaaaaaa aaagtagtta aaagaaaaag atcctaagtt 13440

tgataaatga gttccaaaaa acaaaaagtg gtaagttgat tcacctaact ttatcactt 13500

aggaaaaaat gaagggaggg gaatcctttg acttgctctg cctcccctac tctcttctcc 13560

ccagccttcc ctgctaattc tagctcccct ccaaattcta gctagacctg ctcctccagg 13620

gccacagcag aactgcaaag agaaacagaa gactccaact cttggctttc gcacttccaa 13680

gggtctaagt cttggtgatt tcacagtgag ttaaggaatt gtccatttg gtgaagtgtc 13740

ttcaaaacct aactcccaag ctggatgtta ctgtactcta gccagtgtga aagatctttc 13800

tccctgcttt taactactgg atacaagtaa catctgaggt ggaaatgctc aataaaatat 13860

ataaaataga taaaaagcat ggagagtcct caggcagcct aaatcttcac tgctgggcaa 13920

aaaagcgccc agttagcagc acctggggag cacctttacc aagcaggagg tcctgcgggg 13980

ccagcttgag agccactggc ctggcctccc gcccatctgc tccacgtgcc ctctgcccac 14040

gcagggggag cccaacgtct ttaggcctac cctgattgtc ctgcatgcac atcgggccgt 14100

cacgctggta gttggccact cgagcacggt agggacagtt cacaggtata tgaagataat 14160

tgggtcccag gcgatggcgg tgagtgtcag gataggcaaa aaggcggccc tgcagtaata 14220

aatacaatag acactgacta ggggcagtaa catataacat aggaatgggc tgtaaacttc 14280

attctgaaat ttgaagtaca aagcactctg tacatggtta ccacaaaatt ctgcctttcc 14340

cgctacttaa gagtgaagcc tctactctta tgaaacgagt tctccctaac tgaacagttc 14400

tgaatttccc aggccctctg gccaaccata aatcttcaca tctgtttttg gtaataatat 14460

cttttttttt tccttctgca taacaagtta gtgtcttgtc ttatacagta ttacttccta 14520

ttcccaagga agtattgcat tataaaataa atgtgctaga acgttaacta actgaaatat 14580

tgaattaatt ctattcctcc tcattccccc actccaggga aggaaccaat catttgctct 14640

ttacaaaaag caacacattt ctacaaaacc aacagcatgg gaaattcttc aaataagcac 14700

ttccaactgg gcacagtggc ccatgcctat tataatccca gcactctgga aggccaaggt 14760

gagaggatcg cttgggccca gagtttgag accagcctgg gcaacacagc gagacctgtc 14820

tctacaaaaa aaaaaattaa aacattagcc gagcatggtg gcatgtgcct gtagtcccag 14880

ctactccaac aggctgaagt gggaggactg cttggcccag gggattgagg cagcagtgag 14940

ctgcaatcgt gtcactgcaa tccagcctgg gcaacaaagc aagatcagaa aaaaaaaaaa 15000

aaaaaaaaaa aaagtaccgc ccactcccat gctgccttct gaattatctc agaaacactt 15060

tgagctctga gactttctag atataaacaa caataacaaa aaatctgtgg ttcaaaatga 15120

aagccctaca atgccaaagg taatcaatct aagattaaat tatgttccat gttaatcaag 15180

gcaaagatgg caagcatctt gaaatttctt tcagttaatg ctcaaacaaa agctttatga 15240

taaattctgc attaatctta atatctaatt attgaactat tctcattatt catactcctt 15300

ttctatgagc aatttttctt ccattgaatt tatcattggc cttgaacctt aaaagtattt 15360

ttatatccct aattctatga ggcaaaataa aaaccaaatt gaatagggca acttaagcaa 15420

taactttgtt ccaaatcact gataccaaca gacttaatcc agtcccagtg ttctgtcaat 15480

attttgcttt tggctgaatg actccctctt atcatcattt caaatagcct aaggtcatat 15540

ttagatgact agccatgcgg ccatgaccaa aactcagcta taggttggaa gctggttgtc 15600

tcctttggct caaaaagttg tgctcttgac tccatcaaca ggataggaga tacaatcctg 15660

ccatgggcag ctcctggatg tagcctctat cacctaattc tgattttgtg tcaccctgtg 15720

tttatcaggc aaccttaccc cacctatggg tggagaccaa caaaggttgg gaaatgggca 15780

cagtgaggcc ataccctgac accagcttga atctaggcct tccccaaggc cctgtgcctg 15840

aactcccttg ctggacctgc gtgagtggga aagcctttcc cacccaagca accggcccaa 15900

cttctccaac tcttggcaaa atgcagagaa tgagagcaag aaaggctgca cctgtgtcct 15960

caggtactca ggggacagga cagtctctct atgacttgta ttatattatt tgtccatttc 16020

accatggtgt ggctcaccac tttggataaa gttataaaac ttaataacat gaccttgtta 16080

aaaggcacct gtaaacactg cccattaata tatcaaccgc cactgcctag gctacaatac 16140

tcatgagctc tcagcaggaa taaccccaaa gcccccctaa ctggctttcc acttcctgtc 16200

caacaagaca atctacatat tccctactca cctactcaaa aaatctcaaa tcattcctta 16260

atggtcacca aatggctaaa tgagtacaac agtccttccc caactgtatg gctcctcaca 16320

ggctccatgg tattggaact gatagctaaa tgacctaaga aggatggggg aaagctaggc 16380

tagaaacaat aactcgacaa acttatgaaa gctctttcga acagtttcta aaactcattt 16440

aaaaaaaaaa caacctataa agagtacact gaatttaaaa tagatatatt tgccagaact 16500

aaaattaata ctaacagaaa ttatattact ttctacttgt catccttttt cagaactctc 16560

ttccccactg tttgaatagc tgccttagac atttgctttc ttaggaatgt agagaatacg 16620

tatttgcttt gcagtccttt tcccgggggc ctagttctcc attcaaatct tcattgttct 16680

tcaaattata tgactcagtg ctgctgactg cctcacggtc ccactgtggt tccagagacc 16740

catggtgact tggattcttg tgaaaagagt ttaggtaacc ttgtattcct caatccgttc 16800

cctccctggg taaccagggt tatgcagcct ctgctctgcc catgacaatt tcatgccttg 16860

gcaaatccca ttagcctcat cctctcacag ttcagccacc aggtagggag ttttagggcc 16920

caggattacc ttgttttggc ttcctctact ccctggtttc aatgcaaact tctgcttttc 16980

ctctgagaac tgcctgactt tccttttgct attcccaatc cctagttgtc gctttttcta 17040

tttatcacca agaggagtga ttctactcaa tatatcccag accacacttc tcaaaactga 17100

agaaagcttt tcactcctac tcaagcattt aaggtcccct ctgacatata ctcaatgaac 17160

ctacccaaac tctattcttc tcattcagga gagagtgact gtttatagtt tcaacacacc 17220

taacatgtcc cacctttgtg cttatactca tgtccttctc ctgccgcatt ttacctacta 17280

aaaatcaaaa tcctatctat tcttcaaaac aatctcaaat gctacctcgt tcatgaaact 17340

tcgtggctga cctcctatca gacaaaattt tgctttctgg atcaccatgg cattctgcct 17400

tacattatgg cacccaagac agtccaccat acacaacaga caggtactca atttgtaaca 17460

gttgaattgg actaaaaact aatgaaaaac ttactttcta taaaagcctc agatgttccc 17520

caatgtcata agcaaaaaag ttatttttta aaaacggaat cttagcttgt cctccatgca 17580

gactgaaacc tgaactggaa tcagatactt acataagaag taaccttcag tcgctgtggt 17640

ctgatttagg gtacaatctt taacctttgt ttcaaggtca cagtctaaaa ccacaaaaga 17700

ttgattatta cagtaattat taagccaatt attttcttac tgattctgat ccaaagtaca 17760

atttttcact cttggtactc ttgaaagaga tattaaaaag tgatgtaaaa catactgatg 17820

gcttaacatt agcaaggtga atggttaatc cactgctttt atcactggca gcatacatgt 17880

agttcataaa gatcatccct agatctcttt ttacaaatgg aaaaattcaa gcatcaccat 17940

gaaacagaaa atgaggccag ccacggtggc tcacgcctat aatccagcac tttgggaggc 18000

ggaggcaggc agatcacttg aggtcaggag tttgagacca gcctggccaa cagggtgaaa 18060

ccccgtctct actaaaaata caaaaattag ctgggcatgg tggcgggcgc ctgtagctaa 18120

tcaagaggct gaggcaggag gatcgcttga acctgggagg cggaggttgc agtgagccga 18180

gatcatgcca ctgcactcca gcctgggcga cagagcaaga ctctgtcaca aaaaaaaaaa 18240

aaaaaaaaaa aaaaagaaca agaaaataaa gttaggctta ccaaggtatt ataattaatt 18300

aaccatactt cagctacaga cagaacaaac catgcttcac tctatcaata cttaaggtat 18360

ttctattatg tctttttttt ttaattaaga gagtaagtat tcttgtcttc acatatgtag 18420

ggatcttacc atggactttt gtttatactt ggtttatca aggaatgctc attgaagaat 18480

agttcacctc agtgaatccc acaaggtaac caagcaccca gattaaagtt taagtaatgt 18540

tcttgttaat tcctcacata aagatcatgc caagctgcat attttgtgtg tgcatgctca 18600

catggacacc tgcctcagaa catctcaatc caccaggctc acctgaagca ttttgtcagg 18660

actggcctca atgccaggtg gcatgttgct tgggtcgaag gctatctgtt caacctcagc 18720

aaagtaatta actggattcc ggtttaagac cagtttacca actgggatga gagggtagtc 18780

cttgtgaggc caaacctgaa aatgatgtca tattcacaat taccaggcaa tcaactgaag 18840

gaaatcaaag ctacccacaa aaataccaga tttcaaatac aatcaagcca atgaaaataa 18900

tttttaaatc caataaaaca taagaattta agatgagtgc cttagttaaa tctttcacaa 18960

tagtaagtgg taaatactga cagtttcccc aggaaaaagt acatcagctt ttcataatct 19020

gttccctgtt aataacaata tatgaagcca catacacata ctcaatgaaa tgacaaacag 19080

gaacgtggaa gtaagatgct ggtaagcact cattcacagc ttaggacact ctgaagtggt 19140

accctctgta ttttgtatat gacttcccca ttattataat tgtttgacta gatttttttt 19200

ctaattaggt aagaagagac ttaaaaaaga aaacaatata gttgtttact gactcacctt 19260

ggtgagatcg aatggattaa atggaaaagt ttctgcctga ttaaatgtca tgacctggat 19320

gtaaaaagtc caggaggggt actttcctgt ggcaatggcg ttaaaaagat cccggatgcc 19380

atagtcagga tcttcctggg aaagtctcgc cgcatcttca acagaaaggt ttttgatgcc 19440

ctggtcagtc tacaaaatga aagaaagaag ctccaatgga aacttattgt caaagttatc 19500

aaaatttcat cagtaattca ttgaaggatt atgagtaaca ctgcccaaag aatgaacttt 19560

ctttacaaca atacttctat tgttcaaatc tgatactaat gaatttatca acaagaaatt 19620

ctacttgatt atcccaatac ttttaaatgt caagggaatc ccatctacac atttcatatt 19680

taaaatcagt cctatctttc cctttatgta ttctattatt tgaattttta cactcatgtt 19740

actttccaaa tcagaaaaat caaaatatac ttaaaagtag gcaaaagatt aataaaatta 19800

aaggactact tgccttttat catgaagtaa tgttttaatt ccatttcatg acagctcaaa 19860

tcaattactg agactgtgct cggccagtat cccttcaccc ctgtcctatt cctttttttt 19920

ttttttttaa taaatcatag aatcatagct catagagttg aaaagaactt cagtacagcc 19980

acctggaact agttctctga ctatgagcag gcaaaattaa tattctgact ttccaagcaa 20040

gacattaaaa cccagagaag ctgtgtcact tcataagtgc tcctgttatc atatcatact 20100

gttctttgat ggactgacca tgtgatctta ttctcaaaga cctcccaaca agccaagtgc 20160

cgcagaaaga tttccttagc aaaaatcaac tctgaacgct ggtctatttc taccacagaa 20220

atcccaactg tctcatcagt gaaaaggttt gaactcttac tactgaacaa gaagtaaagc 20280

caccttcagg attaaaacaa tgaaggtcac catgtacaga tcgcccaaag agctaatata 20340

ttaagtagta caataacact gcaatacctt tctgatttta actgaatcct ttaggtttag 20400

aaaagaataa tcttaggtca gaataatcca gactgatgaa tccatgtaga tctcaggcta 20460

ccttagggcc ctaggataaa agacccacaa gcaatccaat ctttccagga cttgtatagc 20520

tggtaatatt atgccaactt cacagtgaaa agaactggag ctcaaagaac ttttaatact 20580

atgcccaagg ccacaattat ctaggagagt agaaatttga gttcaggctg gcataactcc 20640

aaggctacca actagactgg ctgtcctaga tacaaccact tagtcttatt ctaagtaata 20700

ctacattgac ccttttggca attacaagcc aatgggtcaa ctgtttcata aaatttttcc 20760

ttgtcaaata ttatagatcc aagttatgaa tataaaaaga ggtgatagtt ttcctgtttg 20820

atatacctac attaaattgt tcagataaag aagaaagcag gatagaagaa aataggggctt 20880

ttggatgtca ctaagagaga gattatccag tactggtcac gagcccactt ctttctgctt 20940

atcttagcag cagtatttgg aaacagaggt tctcaggctt gtttctgact agcccttggg 21000

agttctctgg gaatatcact aacatgccag ggttcactgg caatattgct cattgtaaca 21060

aatgccatgc ttataccaaa agagggccct ttataatatt tcagttgatt aaaacattct 21120

caaaaccctc cccagatgta actgatttga tctggctata tgaatgcaaa tttttaaaaa 21180

cagtataatt ttgttttctg acatttgtaa ttgattttac ttatatgcct tgttcataat 21240

aaaacctcaa taaatgctaa gtaaatagat ggatgactta aaaagttaaa agaaatgata 21300

ttggaaagac actattggat gttggaagtg ataatgagat tgggatacgc tccaaggtcc 21360

tcagatggta gccaacaaaa tcagtactga tcaatgagat caatacttcc caactttggg 21420

gatggaggtg agggaaaagt gggagaagcc ttaatctttg ctcaatcctt caggaaaaat 21480

aaggaggtat gcggtagtagag ccttgtaccc tctgccccaa aggtaacaca taccttataa 21540

tggaatttgc aataaactgc ctccccattt gcattaacca gcttgaaagt atgtgatcca 21600

tatccattca tgtggcgatg tccatctgga atcccccgat cactgaacaa gaaagaaacc 21660

taaagaacag aaatataaag cctaaaatgt tttcatttaa atcaaaacat ccttgatttt 21720

taagatattc cttagtttcc agaaagtccc ttaatgacat aaaacatact agccaatttc 21780

ccatgggaag ggttaccatt attatttcac ttgtagaaaa taacaaacag aaaaatttta 21840

aatcaaaatg atatgcaaat cactaatact atgactcaaa attctatgaa ggaaagccaa 21900

aacaacctga aaactggcct aatttgttat tcaaaattta tttgcatagc aaatttgttt 21960

ggtactactt tccccccact tccttaatac agagctttac cttacactac agactttcat 22020

ccttactact gctaaatatc aactgacaat gttgctagct atctatcatc atcatcatca 22080

tcatttaagg ccgatgaatg ggggatgatt agtaaagcca gttactacaa aactcttcac 22140

gatcaagaaa tctgaggaag ctgattccaa taaaatagcc aaaactggag agacacttta 22200

tgaagaggca taattaaaca ctgcatccca ggtgatataa tacaatggca aaaaagggtt 22260

catacctgat gcagagactc aggacgtagg ctccagaagt cccagaccat gtccggatcc 22320

ttcagatgtg tctgaggatt tctcttttgg ctgtggataa aagatggaaa ctaaatagaa 22380

aaaagagaga aaaaaatcca aaaactaagt ttacaggaat tatggtgtct ttcatttat 22440

atgcctatta ctaggtttat ggattataat tctaaataca atcctagaat tttaaactgc 22500

caaaccataa tatgaaattt actcagcata gacaactagc atctaagata ccctaaagta 22560

tttcaaatat actcaaaact atagtctttt cttttttttt ttttttaaga cagggtctca 22620

ctctgtcacc caggctgatt gcaatggcat gatcacagct cactgcagct ttgacctcct 22680

gggctcaagc aaatctccca cctcagcctc ccaagtagct gggactatag gcatgagcca 22740

ccaacacttgg ctaatttttg tatttttttg cagagatatg gttttgccat gttgcccagg 22800

ctggtctcga actcatggac tccagtgatc tgcctgcctt ggcctcccaa agtgctggga 22860

tttatagtat ttcttcacgg aaaaaatata cccaccctcc cgcatgcgcg cacgcacaca 22920

caccccctcc tgctgactca gagcatgtag agacaccctc tgcttgtttc caaagtaaag 22980

tccctctttt tactaatctt cgatgctctg caaggaaaac taaactcact tcccattaag 23040

tcaatctgtc ttaaaatgac aaatctaaga ccaatggtaa ctgttcttac tttatcttat 23100

taaatacaga aaaatgtgtt gaaaaaagta gtccagacaa ctcgcattct tttctttctg 23160

cctttcccac ggcttctcct caaatgccaa cctatttgaa atcaatttaa gtcaacaaac 23220

atttgttgag tgcaaaatac tctattacct accaatatgg gatccctgat gaagaaaatg 23280

ggggtgttat ttccaacgag atcccagtta ccatcttctg tgtaaaattt cactgcaaac 23340

ccacgagggt cccgaactgt gtcagctgaa cccgattctc cagctacaat attcaaatgg 23400

aaacaggaag cataagccac agcactttgc atccaagaag cctggagaca taaagaagca 23460

cctggatcca tcctttctaa tccacttcca agaactaaat taggtattta agtgcccggg 23520

ttaattatttc tacccaccag ctaattcagt ccacaaatca aacaaaataa ttaattgaga 23580

gcaataagag aacacatgta ataacagctg acaatgtcat tcatttacca taataagata 23640

actttgttca aacaggtaat tgaactttta attaaaataa gggaagaaaa taattatgag 23700

gcttaaagta gtgacaaaca gaataagtga gttgtcaaaa tgatgaggtt aacattaaga 23760

aaatgagtta aaatattggt tcttagttct taatttcatt tatttttaca ttccacagta 23820

tttttaaaat gcagagactg acacagaaac ggaactagtg aagcactagg tatgttgaaa 23880

tatagtatga tagatctaat actatctctg taacgtgaga aacatcactt agcctctcaa 23940

aaataaaaag aatcccactc tgcaaagtag atctacaaat cagagattat atctgtaaag 24000

cacctaacca cagttggcac agaattaatt ctcaataaag gtcagctgta ggcatagtta 24060

ttaaataaga tttccactct attcttctgt tagaggcaac cacttttagg aaagctcttt 24120

ccaaatgtca cagaattgca gaggtagcct ctgtgtaagt gtgatttcct aagacctcaa 24180

caaacagaaa tacaagtaac ttcctatgtg tctccaaata tgtcttctac tgtaaaacct 24240

tgacataatt tggagaagtg cttgcttcct attttctaat aactttttct ttaagttata 24300

aacatcccca aaaggtgaag ttgagttaag ccataccaat cacgccaata aaccaactta 24360

ccaacagtgg agaaccgaac tgcgatggga gtcttctttc caatatgctc aaataccttt 24420

gccttggagt atttggtaat gtcatgtgtg acctcaaagt agccaaaggc ccctaacgag 24480

aaagacacag aaatccttac catgagacca ttactcagac attcaggtcc tcaacaggca 24540

cctgggtgac aagagaaaat gatggttagc accagccttc tgggagcttc cagtatgttg 24600

gaggaagggg aaatgcccaa acacaaaata aagagggtga aatgcaaagc cctattctat 24660

cattcagtgc taagtcgaat gccacatagc tggagccaag agaaggaggt gagtgaggct 24720

agaagtcgtc aaggttggcc caaggacaca gggtctcaga ggggcctgaa gtacaggtag 24780

gacctatgag agaaagtgaa gaacagagag aagaggagat gaaaacataa gagaaagaat 24840

cttttcaaca cttagcaacg atgatacaaa gcaagcaata aactaaacaa gtcaagatgg 24900

aaaaatacac acaaaccact ggagaagatg gggagaaacc ttatccagga agaagagatg 24960

aagaaatgca gaagtcctaa ctggaagtcc aagtcagcaa gctaagagaa aaacttcctt 25020

tctgtggagc tctttttcca actgagtttt tcactttctt tcaaatacca cccccatctt 25080

ttgtgtctga tataacagcg ataaagaaag ttttacagaa attgttggaa aaaaggaaag 25140

gaaaaatcat cacaaaccca ccaccttaac ataactaccc caaattttgc agctcattcc 25200

agtaattacc cagtaagtac atcatttaaa cttagttgca attgcagtgc atgtgtctat 25260

gtctgtattt ctgtacagca ggtcctcaaa caacattgtt ttgttcaacg ttgtttcatt 25320

ataacgttga tgagaaaaaa atctattcct agctgaggcc accatctgtg tggagtctgc 25380

atgttctccc catgtctaga taggttttct cctggtactc cagcttcccc ccacatccca 25440

aagatgtgcg tgttaggtga actggcatgt ctgaatggtc ccagtgtgag tccaccctgc 25500

aatggacagc atcctgtcca gggctggctc ttgccttgta ccccaacctg ccaggacagg 25560

ctctggcctc ccaccacctt gaactggaat aagctagtaa ataactatct tattttacta 25620

atctttctta aaggtatgta tggctcacat ttatttcaat gtttaatata agggtattt 25680

tgagtctcta tttagaagtt tgatgatgtt ttatcaatca gaaatatgcc ataggaactt 25740

aactcttgct tacgtcaatt agcctatcat aaaattggtt tcattacatg tcgtttaaag 25800

tcaaagtttc caagaaccta ttgatgttaa gtgaggactt attgtatttt tattccttca 25860

atgatggggag aaatggaaga ctgctatttt taaaagaaac tattttatat ataatgttta 25920

acctgaggaa ataaccatca tttatttact aatttcccta ctgctggatg tttttccttt 25980

ctcccacaaa tggagatttg acacttgtgt taaaggtctt ccaatccttt gggaaataac 26040

acttgaccca ggtgctgtga aacaatcttt ttacagcaaa taaacacagc acttacctgc 26100

tcctttagca tgcacaactc tctcaggaat tctctctcgg tcaaaatgag ccatttcatc 26160

agtgaaaacc acatcctgaa caagaagggg cccacggggc cctactgtaa taacattaag 26220

tttgtctcct actgggttac cagctccagt ggtcaggaca tcagctttct acaggaacac 26280

agaaagaaag tgcaggagag ttagcacaat caaacctttc atttacattg tcacgggtac 26340

atagctttgc aatgtttata ccatctgccc tcttttttaa tttcctgtgt ccaaagtact 26400

aaaatctgac aggatgggcc attgcctgaa acttttagtg atgccccaga aacccttgac 26460

aaatctcaac actaatctgc taagaaactt gctctgaaga gcagaaagat gattttggag 26520

cctgaaaatg tgctcagcac ctgctccaga gaaagccact gggttctgca aagctcccag 26580

tcagagaggt gcccaaaagc gtcctacaag cttaacagtg aaatgtccaa tccagcaatg 26640

agaagacggt ctccagggaa gctgtgctgg tcaatacttt actcagaact tacaaacgct 26700

ttaattactgc cacgatcttc aagtgcgcta ccttgtttag tgcatctcaa gaaatgtaca 26760

ccagctattt gcaacatctc ttttcaatta aaaaaaaaaa gtcaatggct ttcactgtca 26820

ctaaaagtaa actgttactg gtaaggtgga ttatagtatt aatgttttaa agatagcaaa 26880

tgtctctgaa atgggatagg ttatttact gtaataaata attaatttta atccacctga 26940

ttctccaatt tcagttacat cagaagacac tcagctgtcc ctcctctagg gttagagcgg 27000

cattcttcta gccccacatg atcctggaag ctcatagctt catctagttc catggtcaga 27060

gaatactttt gaccacagac aactgcctct gcagtgtctt ccccgggtca ctggaaatca 27120

ggagagacac gtaagtgacg caaagctacg ccacctccac ctgggagagg aactcaacgc 27180

tgtctatgac aaaactcaac agaccaactt cacacacaga gaaaagcact atttttagta 27240

tttccacaca cctagagtgg agtctgtagt tttactcccc atgcatgtct gtcctgtaca 27300

ttaaaataaa ttcctgcctc ttctctagaa ggcaatacgg tggggagtag gcacttacaa 27360

aaccacaaaa gatctgtgag ctgtttgaaa aaagatattg aataaaggag accgtgcaat 27420

ttgcaaaact aataaacaag accctctcac acactagtaa atgcccttgc cactagagtt 27480

atatcttcat ccaagtagga agaggccaac atggcagcag ggagaaacta tgagtgccag 27540

cactagagcc tgggtccaac ttccagttct accaattacc agctatgtag ctgctggtaa 27600

gttacttatt tatgagtctc cattttcatc tgcaaaatag aataataagt acctcctaag 27660

gttaacacat gaattaaaag agttaaatgt aaaatgcaag ccacacattc aacacatata 27720

tgctgtcact tcactggctg tgagaacatg acctcgcagc agaaggagca aagcatttga 27780

agttttattt ttctagtaat catgacagat gcatactcct ccatccacac tctctcactc 27840

taccatctct gtccctaaac ccatctagca aagccccttc taaactgtca cagtttaagg 27900

gtccaatcaa cacttaccac ccagtcacat cactcttttg aagatacata tgttgctaat 27960

acgtgtgacc ctctgtgacg tgctaatacg tgttaccctc tgtaggttta tttttaattt 28020

aaagtagaaa atgaaccaga aaaaaaattt ctactttcta aagtagaaaa tgaaccagaa 28080

aaagagcaat ttcaccttta gtttaaacct aaagccttgg ttttcaacct tggcaatgca 28140

ctaaaatcac ctgaggagct ttttaaagcc tcctgatgcc tgggccatcc cccctagaga 28200

tgtctgatag aagtgccctg ggtgaggcct gggctatttt taaatgactc ctgttcaagc 28260

taggatgaa aaccactaac ctagaactat tgggtgataa caaagttaaa ataattagag 28320

agccagagca aaccaaatct gttcactctt taaatgctct tttcaactta caaacaaggt 28380

atttaaggaa gcctttaaaa aataaaggga catccaggct gggcgcggtg gctcacacct 28440

gtaatcccaa cacttcggga ggctgcagca ggtggatcac ctgagatcag gagttcgaga 28500

ccagcctgac caacatggag aaaccctgtc tctactaaaa atacaaaatt agtaccaggt 28560

gcagtggagg ggtgactgta gtcccagcta ctcgggaggc tgaggcagga gaatcgcttg 28620

aacccagcag gcggaggttg cggtgagctg agatcgtgcc attgcactcc agcctgggca 28680

acagcagcga aactccgtct caaaaaaaaa aagggggggga catcccataa aaggtacaag 28740

tatgagaaaa gataccaaga agctcagtaa aaggaacgct aaacagatga gagtgttatt 28800

tacatgccag cattcttaac gtggggaggg agctgcccct aaggcagaga tgagagcaca 28860

tcacatgttc ccttctggtt accgggtcag ggtgatatgg tcaagaataa ataccacacc 28920

caaaggcagc aggtgcagag ttggataaat gaataaagta ctaaagcttg aggtggtctt 28980

tcctgatgac actgactatt aagagtggga atgctggaat tatggcagtc ctccctcctg 29040

cccaatgaag gaatcccctc cttaacatca atcctgttaa gaaatgaaac ctgagttttt 29100

aagttacaaa tggaggcttc agaatctctt ccatttcccc actctaacag ttagatcaat 29160

tgtcctcatt tatataacag ctttccccac aagctcatgt atatacaatt taccttcaaa 29220

gccagtgggg caggaaagct tcaacagtca gcaacagaaa agggaagcca gggagcaaaa 29280

actagaaagg aaaagatccc ccaacttcag gggatgttta gcgcaacatt ggataagagg 29340

aactgatcta aactgaaaga tagagtatta atattagaag gtgaggagca ggatgcggtg 29400

tattagat gaaaataatt aatgcaatga agattacta gatgaaatga gagggtgggg 29460

ctagaaaacg gaccacgcaa gttatctcac ttaagcctgt caataatcca gagatcaata 29520

ttattaatcc catttaatgg gcgatactac aaactcagaa aacttaagta actttgcaga 29580

gatgtcagtg ttgtggcaca ggtttacttg ttacatgtaa cacgtaagga caggcaaaga 29640

gatttcctcc aacttgcaat tattgttttt aactcaatga ataactttag agccagaata 29700

tttttaaaga gagaaaagct tctctatggc tggataattt taagacctct ctctggagag 29760

gtccttgtta acatgtttac atcttcctgg ctgctagctc ccatgacatc catacgtacc 29820

tctcctgaac tgctctctct accctgcccc acaccactca tgaagcaggg ccaagggcat 29880

cttcttgtgc cccatataac aaaatattgt aattgcctgt ttatgtcact gtttttcttt 29940

tttttttttt tttttttttt ttttgagatg gagtctcgct ctattgccca ggctggagtg 30000

cagtggcatg gtctcagctc actgtaacct ccacctcctc ccgggttcaa gcaattctcc 30060

cgtctcagcc tcccaagtag ttgggattac aggcgaccac caccacaccc agctaatttt 30120

tgtattttta gtaaagacag ggttccatca tgttggccag gctgatgatc tcaaactcct 30180

gacctcaaga aatccgccca tcttggcctc ccaaagtgct gggattatag gcgtgagcca 30240

ccgcgcccag cctgtcacta tttttcctct ctagactgtg tactcctttt acatctatat 30300

acctgatccc taatacactg ctcaatatac taacaaacat aaaacaaatg tgtgtggatt 30360

cagcatgcaa ataaactaaa ccacggacca atatgatatg gccaagagcc tttctaacaa 30420

gcaggaggaa gcagcaacag tctatccctc tccctgcatc gcccctccaa aaaatgagag 30480

aaatatttta atttataaat tctgatcaaa cacctttaga cacacttgtt atgaaaactg 30540

ccacagaaat tagcttggct aaaaactagc acaattatta gaagtaggga cttaaagtaa 30600

aattctattt tgactttgct gcctttatag gcttcttttg gccagagcta atacataaat 30660

gaagaagttt tcctgggagg agtaaggttg gcttcatctg gtgaaatatt ttatcaacat 30720

attacacttt ccacttagta gttcccttca ctcacagaag tttcaagaac ctaaacatat 30780

tcttccactt ttaattaaaa gattggttaa tgcaaattgc ccatcagtcc cagaggagtc 30840

ataatgacgt cttttcattc tgtgataaag aatgtgccta gcgttttcta gcaaataact 30900

gcctaacatt ctccattatt tcaacaaaga ttcaagtgcc tacaacgtat acacccacgt 30960

acaggaaacc aaagactgtc ctgaccttca aatttcttgg tctgtgggaa aataataagg 31020

gtggtctaga tattgactgc tgtggaagct aaagcaattc cgtcttagat actgatccac 31080

catgctgact tctgattaac tccagtttta agatttccag tttatctatt attccttgtg 31140

aaagagcccc tatttactat aaatcctgcc ctgaggtcaa acaaccttga catttatcata 31200

cttcagtcat gccatacctc ccttctgaat cgcccttccc tatggtatta agccctgggt 31260

ctgaggggta atggtatggg atccaccatc ttgtctcgcc aacacgcaag acacagacat 31320

ggtttctggt cctaagtccc tactaaacat ttctttctga gaaacttgat ttgtcagcct 31380

ctttctttgg cctctcagct tcctcagact ttgggggtag gtttgcatag acctacccac 31440

catgcaaaag ctaaaaaaaa gaaatgaaac cttaaaaatt tctgtcccat ctcccatagt 31500

ctctcccact gcacataatg acttagcgat tccataccta ggtatatttc aggagaaatg 31560

aaagcacatg tggaccaagg gttaagtcac tcagtcaccc cttctccatc ccaaatgtta 31620

gcagtgtaac atcaatgcag ttattaccag gattacccta aaaaaatata gaccctcaca 31680

ggctataatg cataggcaac tcactgacag aaatggatag tctaagactt aacctctctc 31740

ccaggtaata agagaactgt ataaaaaggt aaggtttgaa atgttcaaat tctcccattt 31800

taaagggatc agaggctact tattaacatt tccccattatac ccccaagaaa agagcactgg 31860

cagcaagctg cacttaaaaa aaaaaaaagg aaatgggtta aaggtaagta tctcaaaaga 31920

agagcccatg gcattctaag cagtgagcaa taaagttccc atttaagttc ccttcaaagt 31980

ggctgggaat acctgcacaa ccacctccct cctgccaaga gagaaaaaga aataagaatt 32040

tttaagtgga tcgaggtcat actcatgaac acatctgact caatgctcac ttattaaagt 32100

ttacaaaact cccaattcac ctgtcagcaa accaaatctg accacagaag atacgcccaa 32160

taaattgctt tcacatggtt gactgtgttt ccaaaatagg acactctgat tcttgagaga 32220

ggaggcaaca gagtgttcgc ctatagtagt cccaaaccat aaagctctcc tagatcagta 32280

aagcatcaca actaattggg aagtggaaga catcatatta gcacaaagct gcccacttag 32340

ataaaagcat ttaccaagca aggggagatt tttttttaat accatctgtt actgtcataa 32400

ttttctaaca gaatatctta atgtcagaaa gtaaaaatga catattcttt aaaaagaaat 32460

tatgttgcat gtccattgcc gcatcacaag aaaatggctg gacagatttt caccaaatat 32520

gaaatgtacg ctttggataa tatagtagag tcagcttcct ggacaggggg tagactaccc 32580

caggtgtggc ctgtaacaga gttctagcca acagaatgta agtggcagtg accctggacc 32640

ctaaaccttc ccacactcat atacccttct ccacgctctt gagccttctg tgctggacag 32700

tgacgcccat aaaagccatg tttgaccaag ggggagcttc cttcagcctg gtgccctcaa 32760

taactgtggg aagcagagca cccacccacc tccttcaacc tcaaaccact ttgggccatt 32820

catgtaggaa aggaatacat ttcttttttt ttctttttga gatggagttt cactcttgtt 32880

gcccaggctg gagtgcaatg gcaggatctc ggcttactgc aacctccgcc tccagggttc 32940

aagtgattat cctgcctcag cctcccaagc agctgggatt acaggcatgt gccaccacgc 33000

ccagctaatt ttgtattttt agtagagaca gggtttctcc atattggtca ggctagtttt 33060

gaactcctga cctcaggtga tccgcccacc tcagcctccc aaagtgctgg attaggtg 33120

tgagtcactg cacccggcca taaatttcta atatgtttat accatgatac atgttgagat 33180

ttattattat aacacttagc ctacacatga ttccacatga ttgaacttaa aatatgggct 33240

atgaattaat aatttaaaag caattcaatt tttaaaaaga cagaaaattt gaacatatac 33300

ttcaaaggaa tatataccaa tggtcaataa gcatgtaaaa agatgctttg catcattagt 33360

catcagagca aagcaaatta aaaccataat gagataccac tgcacaccca acaactaaat 33420

gtaaaagact gacaacacca aaagttggta ggatgtggag caactggaac tcccataacac 33480

tgctgatgaa gtataaaatg gtacaaccac tttggaaaaa ggctcagcag tttctcataa 33540

cacattcact taccttatga cccaataaat ctactcctag gtatttccta agagaaataa 33600

aaccacctgt gcacaaaaag acttatgtaa gaatttttat agcagcttta tttatttta 33660

atttttaaga cagggtctcc ctctgtcacc caggctagag tgcagtcatg taatcatggc 33720

tcactgcagc ctccacaccc caagctcaag caatcctccc aactcaactt cccgagtagc 33780

tgggactaca ggcatgtgtc accatgtctg gctacttctt ttatttgttg tagagatggg 33840

ttttcaccat gttgcttagg ctggtctcaa ctcctgagct caaactatcc tcccacctca 33900

gcctccaaaa gtggtagggt tataggcata agccttcatg cccagccaca gcagctttac 33960

ttacaataga aaaaaaatta aatagcccag gtgtccagca acaggatatc caaatggtgg 34020

tatattcaaa taatggaatc ttactcagca atgaaaagaa caaactgcta atacatgaaa 34080

caacttgaat gaattaaagg atggacagac agatggagta acgtaataaa aagacacaggg 34140

caaaatgttc cttgtagaat ctagatggca gatatataga tatacactgc aaaatttttt 34200

caacttttct gaatatttga aatttttcat ttaaaaaaag tgtttgaaaa ttatataggc 34260

tataaggcat atgcataatt ccttttggag ggccctgggt ggagggagaa gtgcctaaaa 34320

gaagtaagcg gtttttacta agagcagcta aaactgagga acagtaagac atcaatgtga 34380

ctgccaatca ggacgatgta cacagtgccc tgagatgtca tggacaagg ggatggctgt 34440

atttcaatta agagcctccc aactatagat gtgttttgga gtcagccatt tctcacatgg 34500

gcaactatgt aaggtaaatc aacagcaggg acttcaacag ttcatgattc tcccaaacaa 34560

ggaagaaggc agctagtctc agaataaagg gagtccttt aaaaatgaaa atttacacc 34620

tgtaatctcg gtactttggg aggccaaggc gggcagatct cctgaggtca ggagttccag 34680

tccagcctgg ccaacatggt gaaaccccat ctctactaaa aatacaaaaa ttagctgggt 34740

gtggtggtgg acacctgtaa tcccagctac tcgggaggct gaggcaggag aattgcttga 34800

atccgggagg cggagtttgc agtgagtgga gatcatacca ctgcactcca gcctgggcaa 34860

cagagcaaga ctccgtctca aaaaaaaaaa aagaaaagaa aagaaaaaga aaaagaaaat 34920

attttgtttt gtaaaaatct cactactttg tccacacttt tctcaattca ccgagtattc 34980

atgaaaactt cagtgtgaac caggatttgc accactggg taaggaaaga gtcttgggtc 35040

caagaagctg aattctaacc ttgtctctgc cagtaagtag tggcacatcc tttactaaga 35100

ctctacctct ctctggttta ctttaagagc tcatgaacca acctaatggt atataacaaa 35160

gaacagggcc aagaggggag attttccagg cccttggagg agaatggcag ccttttccca 35220

aacagggaac cattaaagca atccatgagc acttagcatt aataccccag ttcacgaacc 35280

tgagttttga gcaagagcac taggcagaat agggaaaaca gggctgcagt cctctgtcta 35340

ccattaacca gcagccaggg aacagggcaa gacctttaga ctccctgggc caatctgcca 35400

acagtagact cagaccagat gtggtctcat ctaatccaac ctcctcatag atgccctctg 35460

agactctgca gccgtgggca caggccagga atgcctcccc caccaccaag ttaaaatcct 35520

actcatttaa aaaaattcct gttcaaatgt cacctgcttt gtccggccca cctggtcact 35580

ccctctacat tctacagata acctctatta gagccctcac cacgctggag tgtaatcatc 35640

tgtttatgtg tctcgtctcc cagactagac tctcagcttc tcaagggtgt tgttcttgcc 35700

gcactggaat ttgtacacct tgccccgaac agtgcctagc acctacaggt gctcaaagaa 35760

tagaataaac gcattaatca atccctaatg tcacccccca gctctaaaat gctaggcttg 35820

acattttgaa gtgcaaatga gacaatgaat gtaaaagttc ttttatttag tcgtttaata 35880

atgtgcatag tacgtaccag tcactgtgac tggtgctgga gacctcaaga ttgtaaaaca 35940

caatccctgc cctgctggga ccccacaact atgaaagcgt gtaaacagta tttcaagaag 36000

ggcaccatta gagcctggtt ccacttctcc cctcactgac ttccttgcag ctgcagctct 36060

ctgcctcttt ccccatgagc cctcaatctg tcctcaagct gcgacctctg tcccagttgg 36120

caaaagtgca aaattctgaa gcaaccaccc gagcctgtcg cggacaacaa agcctgcagt 36180

cccgcgcccc actacccgga cccccctccc ctgcttcggc gaatgtaaaa gtccgtctgc 36240

accgaagggg atcccccctg cccagtgagc cgcggtacag tccctccaag cgccgccccg 36300

ggggaagccg ggcctttact tgccgacgcc cccgctttct aaacggacct tcgggcccgc 36360

tcggggagca cagagtgtac ctgcgcggcc cgctgctcct tccagtgctg catctggtcg 36420

ctggcgggat cccggctgtc agccatagcg tgcggtttgc tgtgcagaac actgcaggag 36480

gcctcggctc gtgggtctcc accctcagca ggcaaatctg cctgttgccc cgagtccgtg 36540

gcgacttcag gctcagccaa tcagcaccac cccctcgtcc cgccccctcg ggaggactgc 36600

cttctgattg gctgctcggc gctaggcagg ccaagattgg aagcccaatg ggctcggagc 36660

agggctctcc gaaggaggcc tgaagaccgg agatacccag gagggtgcgg aaaggaaggg 36720

tcccgctctg gcccagcaat tggagagcct cgccccgccg gcccggggtg cccgggatag 36780

ccgaacccag ggcggggctc ccggttatca gcatccttca ggccgccctg ggacgcggcc 36840

gttggtggcc tgccagtttt gcagacggaa tacttagacc ctgctgggtg gggccttgcg 36900

ctccatagct atgctttctc agctcttaaa aagtcagcaa tcctgggtta aattactctg 36960

tacaaaatta aattttgcca tacgctaatg taagtattat ttgcaaccaa aggatggatg 37020

gatgcttttc tttttctttc ttagctcttc ttaacttatt cgtggattat cacttactag 37080

actagtatac taattattta atataaaaac actctgaaac gtttatctcc gtacagatgg 37140

gtgttgattt cctcctttac cagagaaaaa caacttaact tagatgggaa aaatgtattc 37200

ccttgtgatg ggaaaaatgt attcaactct ggtacactgc caaattggct tctttaaaca 37260

ctggagaaat ctgcttcccc aaattttacc cccaggtaag caggaatttt gaaaagcttt 37320

gtgtctttaa aaagctatgt tgaagagtaa aatacttagt aatatatagt acaatattta 37380

gtaagttact gactagtttg tcattgtaat caaagagttt gaatgatttc tacacctcct 37440

caggtgctag gatttatctt taagcacctt aatatatata catagtatat accatcatct 37500

tcaaattaaa agttatcagc ttttaatttt ttccagtaca actacatata atgggtggat 37560

atataatagt ggaaacttca catgttgttc ttgatcttac aaaagtatga taagaatctt 37620

gggcataaaa gtggaacttt agcagttcat tggaaaagaa agccacaaat tgatctttag 37680

caccctatac ttttcctata ttaaaaacaa ttataaagat gctgttggct gtgtaatttg 37740

gtgtcttttt ctacctgccc gatccatact gccaagactg taaactccat gtgggtcagg 37800

atcctcccat ccataattta ttgagcactt actatatacc agggactatt ctcagctttt 37860

tgtatatttg cttagttaac cctcccaaca accctatgag ttagatacta atattatctt 37920

cattttacag attaggaaac agaggtatag ggaggttaaa taacttgccc aaagttacac 37980

agctattaag ggatagtcag gattctagct tcagctgttc atctccaaag tccacagttt 38040

gttgaataaa taaatgttga aataatcgag caaatcatct tcttaactaa ccaacacaaa 38100

<210>2

<211>20

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>2

gcatgcacaa ctctctcagg 20

<210>3

<211>20

<212>DNA

<213> Artificial sequence

<220>

<221>misc_feature

<223> Primer

<400>3

ggtttctggg gcatcactaa 20

Claims (10)

1. an in vitro detection sample whether there is a method for the single nucleotide polymorphism of catalase gene CAT, it is characterized in that, comprises steps: (a) amplifying the CAT gene of the sample with CAT gene-specific primers to obtain an amplified product; and (b) Detect whether the following single nucleotide polymorphisms exist in the amplification product: 26328 bits T→C; Wherein, the nucleotide position numbers are based on SEQ ID NO:1. 2. The method of claim 1, wherein the gene-specific primer has the sequences of SEQ ID NO: 2 and 3. 3. The method according to claim 1, wherein the length of the amplified product is 100-3000bp, and contains the 26328th position in SEQ ID NO:1. 4. A test kit for detecting hypertension, characterized in that it includes primers for specific amplification of CAT genes or transcripts, and the amplified length of the primers is 100-3000bp, and contains SEQ ID NO: 1 Amplification product at position 26328. 5. The kit of claim 4, further comprising reagents selected from the group consisting of: (a) a probe that binds to the mutation at position 26328 in SEQ ID NO: 1; (b) Recognition of the mutant restriction enzyme at position 26328 in SEQ ID NO:1. 6. test kit as claimed in claim 5, is characterized in that, described mutation is following single nucleotide polymorphism: 26328 bits T→C; Wherein, the nucleotide position numbers are based on SEQ ID NO:1. 7. test kit as claimed in claim 4, is characterized in that, described primer has the sequence of SEQ ID NO:2 and 3. 8. A method for diagnosing an individual's susceptibility to hypertension, characterized in that it comprises the steps of: detecting the individual's CAT gene, transcript and/or protein, and comparing it with normal CAT gene, transcript and/or protein, A difference indicates that the individual is more likely to have high blood pressure than the normal population. 9. The method according to claim 8, characterized in that, the gene or transcript of CAT is detected, and the difference is compared with the normal CAT nucleotide sequence. 10. The method of claim 9, wherein said difference is the following single nucleotide polymorphism: 26328 bits T→C; Wherein, the nucleotide position numbers are based on SEQ ID NO:1.