CN1661081A - Relativity between gene of catalase 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 disclosed.

Description

The dependency of catalase gene and essential hypertension

Technical field

The present invention relates to molecular biology and medical field.Relate more specifically to catalase gene (catalase, single nucleotide polymorphism CAT) (single nucleotide polymorphism, SNP) and with the dependency of essential hypertension.The invention still further relates to the method and the test kit that detect these SNP.

Background technology

Hypertension is meant one group of clinical symptom grouping that systolic pressure or diastolic pressure raise.The rising of blood pressure and coronary heart disease, renal tubal dysfunction, hypertensive heart disease and the concurrent cerebral apoplexy of hypertension have an apparent causal connection.The up-to-date Case definition of hypertension is systolic pressure 〉=19kpa (140mmHg) or diastolic pressure 〉=12kpa (90mmHg), and meeting wherein, a person can be diagnosed as hypertension.

Most hyperpietic only has slight subjective symptoms in early days in elevation of blood pressure, as headache, dizziness, insomnia, tinnitus, fidgety, the difficult concentrated fatigue etc. that also occurs easily of working and learning energy.Along with the development of the state of an illness, when particularly developing complications, symptom increases and obviously gradually, as numbness of the fingers and stiff, lower limb pain appears when walking more, or the nervous sense of nape portion sore muscle appears.Show when, shortness of breath nervous, uncomfortable in chest, precordialgia that heart gets involved, frequent micturition at night, diuresis occur, show when urine is light that kidney is got involved, the kidney arteriole hardens when occurring.If obnubilation appears suddenly in the hyperpietic, breathe promptings such as dull irregular, gatism hematencephalon may take place, if engender a side limb activity inconvenience, numbness even benumb, should suspect the formation whether cerebral thrombosis is arranged.

The early stage Non Apparent Abnormality sign of hypertension occurs.When appearring in vitals such as brain, the heart, kidney, the mild damage can have unusual sign to occur.The performance of common heart abnormality has that apex beat moves to left, pareordia is praised the sample pulsatory feeling, auscultation apical region of heart first heart sound strengthens, second sound of aortic area strengthens and systolic murmur and diastolic murmur are arranged, show arteriosclerosis and left ventricular hypertrophy take place, if listen in the apical region of heart and the galloping horse sample rhythm of the heart may show the appearance of central force depletion.Also common in addition ear-lobe folding line, capillary pulsation, radial artery occur and pulsus durus or acrotism and lower limb intermittent claudication etc. occur.

In addition, because the development of some risk factor or hypertension itself can cause the remarkable or hurried rising of some hyperpietic's blood pressures, simultaneously with vitals functional lesions such as brain, the heart, kidney, retinas, a series of clinical special signs appear in serious threat to life, are called hypertensive emergency.The sickness rate of hypertensive emergency accounts for 5% of Hypertensive Population, and common have hypertensive encephalopathy, hematencephalon, acute left heart failure, the acute withdrawal syndrome of clonidine, Acute Myocardial Infarction, a radical type malignant hypertension etc.

About 5% left and right sides no conscious sympton among the hyperpietic does not know when blood pressure raises, and does not more know to have produced when the complication of blood vessel and organ injury yet, some patient even just know after cardiovascular accident has taken place and oneself suffer from hypertension.So, find out the genetic cause of hypertension incidence, hypertensive morbidity can be effectively controlled in preventing and detecting early, and disease is reduced to minimum to the injury of human body.

(essential hypertension EH) also is essential hypertension to essential hypertension, is a kind of independently disease, and the cause of disease of oneself, rule and the clinical manifestation that the generation development lapses to are arranged.Mainly show as the rising of arteriotony clinically.Account for more than 90% of crowd hyperpietic, at present pathogeny is not clear fully as yet, mainly just can be diagnosed as essential hypertension (essential hypertension) after having got rid of the hypertension that other diseases causes.The rising of arteriotony mainly is because of due to periphery arteriole resistance increases, and in various degree Q volume of blood and kinemic increase are arranged simultaneously.Often cause late period internal organs such as the heart, brain, kidney to be got involved severe complications such as hypertensive heart disease, heart failure, renal tubal dysfunction, hematencephalon take place.The treatment of essential hypertension mainly is to bring high blood pressure down to prevent the generation of complication simultaneously.The primary hypertension patient cause of death is cerebrovascular accident, cardiovascular accident and renal insufficiency, and for seeing, in heart failure and uremia is taken second place more with cerebrovascular accident in China, and American-European countries sees with heart failure more, and cerebrovascular accident and uremia are taken second place.

Essential hypertension is as modal cardiovascular disorder, and its sickness rate rises year by year, and can cause the serious heart, brain, kidney complication, is the primary hazard factor of cerebral apoplexy and coronary heart disease.EH is a kind ofly interacted and the polygenic disease of morbidity by inherited genetic factors and environmental factors, and the genetic mechanism of seeking the EH genes involved and then illustrating hypertension incidence has become the focus of present research.

Though have many researchs about range gene polymorphism and essential hypertension, do not confirm the report of CAT gene and essential hypertension dependency, more do not confirm the SNP of CAT gene and the report of essential hypertension dependency.

In sum, for the final treatment hypertension that realizes, this area presses for seeks the essential hypertension tumor susceptibility gene, and develops method, test kit that detects essential hypertension and relevant medicine.

Summary of the invention

Purpose of the present invention just provides a kind of diagnosis (especially early diagnosis) hypertensive method and detection kit.

Another object of the present invention provides the hypertensive method of a kind of new treatment.

A further object of the present invention provides the hypertensive pharmaceutical composition of a kind of treatment.

In a first aspect of the present invention, a kind of method that the hypertension susceptibility of individuality is diagnosed is provided, it comprises step:

Detect this individual CAT gene, transcript and/or albumen, and compare with normal CAT gene, transcript and/or albumen,

There are differences and just show that this individuality suffers from hypertensive possibility and be higher than normal population.

In another preference, what detect in the described method is gene or the transcript of CAT, and with normal CAT nucleotide sequence comparing difference.

In another preference, described difference is following single nucleotide polymorphism:

18934 T → C;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

In a second aspect of the present invention, the single nucleotide polymorphism enzyme method that provides a kind of test sample whether to have catalase gene CAT comprises step:

(a) with the CAT gene of CAT gene-specific primer amplification sample, obtain amplified production; With

(b) detect whether there is following single nucleotide polymorphism in the amplified production:

18934 T → C;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

In another preference, described gene-specific primer has the sequence of SEQ ID NO:2 and 3.

In another preference, the length of described amplified production is 100-3000bp, and contains among the SEQ IDNO:1 the 18934th.

In a third aspect of the present invention, provide a kind of detection hypertensive test kit, it comprises the primer of specific amplification CAT gene or transcript, more preferably, it is 100-3000bp that described primer amplification goes out length, and contains among the SEQ ID NO:1 the 18934th amplified production.

In another preference, described test kit also contains the reagent that is selected from down group:

(a) with SEQ ID NO:1 in the 18934th sudden change bonded probe;

(b) the 18934th sudden change restriction enzyme among the identification SEQ ID NO:1.

In another preference, described sudden change is selected from following single nucleotide polymorphism:

18934 T → C;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

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

Embodiment

The inventor is through deeply and extensive studies, and the SNP of a large amount of candidate genes is measured and analyzes.Find first and proved that the part SNP and the hypertension of CAT gene are closely related, and found its new function: the change of CAT will cause hypertension, wherein the association study result shows, (18934 T → C) there is significant difference (P＜0.05) in the distribution in case and control group, therefore can be used as to detect hypertensive specificity SNP at the SNP of the 18934th of CAT.Finished the present invention on this basis.

The CAT gene

(catalase, detailed sequence CAT) can be the nucleotide sequence (can referring to network address http://www.ncbi.nlm.nih.gov/) of AL035079 referring to accession number to catalase gene, shown in SEQ ID NO:1.

Catalase (CAT) gene belongs to more clearly peroxidase on the 26S Proteasome Structure and Function.This gene of people's separation of human such as Quan in 1986 has also been set forth the feature (Quan et al.NucleicAcids Res.14:5321-5335,1986.) of this gene.Full length gene 34Kb is made up of 13 exons.Gene product is made up of four subunits of 60Kd, and each subunit comprises a protoferriheme group, and molecular weight is 240Kd.Catalase can be deleterious H ₂O ₂Resolve into H ₂O and O ₂Make cell avoid the murder by poisoning of oxidative pressure.This albumen is separated obtaining from the red corpuscle of mouse the earliest, also separates having obtained this albumen subsequently at the intravital red corpuscle of people.Catalase and SOD and glutathione reductase have been formed intracellular main antioxidant system.

Catalase is the proteolytic enzyme that has the intravital extensive existence of present known biology, and at liver, it is higher to express abundance in nephrocyte and the red corpuscle.Hydrogen peroxide zymoprotein and SOD and glutathione reductase have been formed intracellular antioxidant system jointly, have brought into play main antioxygenation at red corpuscle hydrogen peroxide enzyme.There are some researches show that Japanese acatalasia and the hydrogen peroxide enzyme concn in the red corpuscle are crossed low relevant, catalatic concentration has only normal people's 0.2-4% (Ogata, M.Aatalasemia. in this disease red corpuscle Hum.Genet.86:331-340; 1991), and be because (the 5th bases G of the 4th intron → A) that montage sudden change causes.(21, A → T, translation initiation site are 1 to find in the upstream regulatory region territory there be polymorphic the existence among the Hungarian; (Goth.L et al.Electrophoresis.1997jun:18 (7): 1105-8).By the end of about catalase gene polymorphic jointly that everybody approved being arranged seldom also so far, what major part was found is have national characteristics polymorphic.

Years of researches show that the unbalance meeting of oxidative pressure causes (Jose M et al., Biochemistry, 32 (8), 595-603,1999) such as the generation of disease such as atherosclerosis etc. and some aging diseases.As antioxidase main in the red corpuscle, catalase has been brought into play very big effect (Gaetani GF, et al., Blood 87:1595,1996) in antioxygenation.

Goth L in 2000 and Eaton JW find the hydrogen peroxide enzyme defect can cause higher diabetes generation (Goth et al, Lancet 2,000 25; 356 (9244): 1820-1).

The deleterious H because the hydrogen peroxide endonuclease capable is degraded ₂O ₂Avoid pair cell to cause the harm of oxidative pressure, therefore the candidate gene as essential hypertension carries out examination at the isolated population of China.

The inventor checks order to the almost whole zone in the CAT gene, has found many SNP, and wherein most of SNP and hypertension susceptibility are also uncorrelated, yet association study shows 18934 T → C but are and the very high SNP of hypertension susceptibility cognation.This SNP is arranged in No. 7 intron of CAT, and prompting is influential to the montage process after the transcribing of CAT, and then causes influencing catalatic expression, and the hypertension susceptibility that finally causes the carrier is apparently higher than normal population.

Based on new discovery of the present invention, CAT albumen or polypeptide have many-sided new purposes.These purposes include, but is not limited to: the direct disease (as hypertension) due to the low or forfeiture and be used to screen the material that promotes the CAT protein function as pharmacological agent CAT protein function, and as antibody, polypeptide or other part.The peptide molecule that can stimulate people CAT protein function that can be used for seeking therapeutic value with the recombinant human CAT protein screening peptide library of expressing.

On the other hand, the present invention also comprises people CAT DNA or the polypeptide of its fragment coding has specific polyclonal antibody and monoclonal antibody, especially monoclonal antibody.Here, " specificity " is meant that antibody capable is incorporated into people CAT gene product or fragment.Preferably, refer to that those can combine with people CAT gene product or fragment but nonrecognition and be incorporated into the antibody of other irrelevant antigen molecule.Among the present invention antibody comprise those can in conjunction with and suppress the proteic molecule of people CAT, comprise that also those do not influence the antibody of people CAT protein function.

The present invention not only comprises complete mono-clonal or polyclonal antibody, but also comprises having immunocompetent antibody fragment, as Fab ' or (Fab) ₂Fragment; Heavy chain of antibody; Light chain of antibody; Genetically engineered strand Fv molecule; Or chimeric antibody.

Antibody of the present invention can be prepared by the known various technology of those skilled in that art.For example, the people CAT gene product of purifying or its have antigenic fragment, can be applied to animal to induce the generation of polyclonal antibody.Similarly, expressing human CAT albumen or its have antigenic segmental cell and can be used to immune animal and produce antibody.Multiple adjuvant can be used for the enhancing immunity reaction, includes but not limited to freund's adjuvant etc.

Antibody of the present invention also can be monoclonal antibody.This type of monoclonal antibody can utilize hybridoma technology to prepare.Antibody of the present invention comprises the antibody that can block people CAT protein function and the antibody that does not influence people CAT protein function.Each antibody-like of the present invention can utilize the fragment or the functional zone of people CAT gene product, obtains by the routine immunization technology.These fragments or functional zone can utilize recombinant methods or utilize Peptide synthesizer synthetic.Can come immune animal and produce with the gene product of producing in the prokaryotic cell prokaryocyte (for example E.Coli) with the unmodified form bonded antibody of people CAT gene product; With posttranslational modification form bonded antibody (as the albumen or the polypeptide of glycosylation or phosphorylation), can come immune animal and obtain with the gene product that produces in the eukaryotic cell (for example yeast or insect cell).

The proteic antibody of anti-people CAT can be used in the immunohistochemistry technology, detect people CAT in the biopsy specimen proteic what and/or whether suddenly change.A kind of preferred anti-CAT antibody is the antibody of the normal CAT of nonrecognition CAT but identification suddenlys change, perhaps discerns normal CAT but the antibody of nonrecognition sudden change CAT.Utilize these antibody, the hypertension susceptibility that can carry out protein level easily detects.

Utilize CAT albumen of the present invention,, can filter out with CAT albumen interactional material takes place, as inhibitor, agonist or antagonist etc. by various conventional screening methods.

CAT albumen of the present invention and antibody thereof, inhibitor, agonist, antagonist etc. when using (administration) in treatment, can provide different effects.Usually, can these materials are formulated in nontoxic, inert and the pharmaceutically acceptable aqueous carrier medium, wherein pH is about 5-8 usually, and preferably pH is about 6-8, although the pH value can be with being changed to some extent by preparation Substance Properties and illness to be treated.The pharmaceutical composition for preparing can carry out administration by conventional route, comprising (but being not limited to): intramuscular, intravenously or subcutaneous administration.

Normal CAT polypeptide can be directly used in disease treatment, for example, is used for hypertension therapeutic.When using CAT albumen of the present invention, also can use the hypertensive medicament of other treatment simultaneously.

The present invention also provides a kind of pharmaceutical composition, and it contains CAT albumen of the present invention and the pharmaceutically acceptable carrier or the vehicle of safe and effective amount.This class carrier comprises (but being not limited to): salt solution, damping fluid, glucose, water, glycerine, ethanol and combination thereof.Pharmaceutical preparation should be complementary with administering mode.Pharmaceutical composition of the present invention can be made into the injection form, for example is prepared by ordinary method with the physiological saline or the aqueous solution that contains glucose and other assistant agents.Pharmaceutical composition such as tablet and capsule can be prepared by ordinary method.Pharmaceutical composition such as injection, solution, tablet and capsule should be made under aseptic condition.The dosage of activeconstituents is the treatment significant quantity, for example every day about 0.1 microgram/kg body weight-Yue 10 mg/kg body weight.In addition, polypeptide of the present invention also can use with the other treatment agent.

When making pharmaceutical composition, be that the CAT albumen of safe and effective amount or its antagonist, agonist are applied to Mammals, wherein this safe and effective amount is usually at least about 0.1 microgram/kg body weight, and in most of the cases be no more than about 10 mg/kg body weight, preferably this dosage is about 0.1 microgram/kg body weight-Yue 100 micrograms/kg body weight.Certainly, concrete dosage also should be considered factors such as route of administration, patient health situation, and these all are within the skilled practitioners skill.

The proteic polynucleotide of people CAT also can be used for multiple therapeutic purpose.Gene therapy technology can be used for treating because cell proliferation, growth or the metabolic disturbance due to the proteic expression of CAT of the proteic nothing expression of CAT or unusual/non-activity.The method that structure carries the recombinant viral vector of CAT gene is found in existing document (Sambrook, et al.).Recombinant human CAT gene can be packaged in the liposome in addition, and then is transferred in the cell.

Polynucleotide imports tissue or intracellular method comprises: directly be injected into polynucleotide in the in-vivo tissue; Or external by carrier (as virus, phage or plasmid etc.) earlier with the polynucleotide transfered cell in, again cell is transplanted in the body etc.

The invention still further relates to the diagnostic testing process of quantitative and detection and localization people CAT protein level.These tests are known in the art, and comprise ELISA etc.

Whether having the proteic method of CAT in a kind of detection test sample is to utilize the proteic specific antibody of CAT to detect, and it comprises: sample is contacted with the CAT protein specific antibody; Observe whether form antibody complex, formed antibody complex and just represented to exist in the sample CAT albumen.

The proteic polynucleotide of CAT can be used for the diagnosis and the treatment of CAT protein related diseases.Aspect diagnosis, the proteic polynucleotide of CAT can be used for detecting the proteic expression of CAT CAT abnormal exprssion whether or under morbid state.Can be used for the hybridization of biopsy specimen to judge the proteic abnormal expression of CAT as the CAT-DNA sequence.Hybridization technique comprises the Southern blotting, Northern blotting, in situ hybridization etc.These technological methods all are disclosed mature technologies, and relevant test kit all can obtain from commercial channels.Part or all of polynucleotide of the present invention can be used as probe stationary on microarray (microarray) or DNA chip (being called " gene chip " again), is used for analyzing the differential expression analysis and the gene diagnosis of tissue gene.Carry out RNA-polymerase chain reaction (RT-PCR) amplification in vitro with the special primer of CAT albumen and also can detect the proteic transcription product of CAT.

The sudden change that detects the CAT gene also can be used for diagnosing hypertension.Detection can be at cDNA, also can be at genomic dna.The form of CAT protein mutation comprises that the point mutation compared with normal wild type CAT dna sequence dna, transposition, disappearance, reorganization and other are any unusual etc.Available existing technology such as Southern blotting, dna sequence analysis, PCR and in situ hybridization detect sudden change.In addition, sudden change might influence proteic expression, therefore can judge indirectly that with Northern blotting, Western blotting gene has or not sudden change.

The method of the detection SNP of the present invention of most convenient is by the CAT gene with CAT gene-specific primer amplification sample, obtains amplified production; Detect then and whether have following single nucleotide polymorphism in the amplified production: 18934 T → C, wherein, nucleotide position is numbered based on SEQ ID NO:1.

Should understand, after the present invention has disclosed the SNP and hypertensive dependency of CAT gene first, but those skilled in the art can design the amplified production that specific amplification goes out to contain this SNP position easily, determine whether to exist 18934 T → C by methods such as order-checkings then.Usually, the length of primer is 15-50bp, preferably is 20-30bp.Though the complete complementation of primer and template sequence is preferred, one skilled in the art will appreciate that at primer and template to have under the situation of certain not complementary (especially 5 of primer ' end) also can increase specifically (promptly only amplifying required fragment).The method that contains the test kit of these primers and use these primers is all within the scope of the invention, as long as the amplified production that this primer amplification goes out contains the correspondence position of SNP of the present invention.A kind of preferred primer is to having the sequence of SEQ ID NO:2 and 3.

Though the length of amplified production is not particularly limited, the length of amplified production is 100-3000bp usually, preferably is 150-2000bp, more preferably is 200-1000bp.These amplified productions all should contain among the SEQ ID NO:1 the 18934th.

Because SNP of the present invention and hypertension have very high cognation, therefore not only can be used in early days diagnosing primary hypertension more exactly, and can make the carrier just take reasonable precautions (as on diet, taking corresponding control) against a rainy day at premorbid not, thereby improve carrier's lifetime and life quality, therefore have earth shaking using value and social benefit.

Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to people such as normal condition such as Sambrook, molecular cloning: laboratory manual (New York:ColdSpring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.

Embodiment 1

1.1 research object

Because blood pressure is the quantitative character that is subjected to multiple such environmental effects that polygene participates in, the inevitable problem that when analyzing genes involved, will consider genetic heterogeneity.Select the relatively single isolated crowd of genetic background will help to reduce the influence of genetic heterogeneity, simultaneously because these colonies have comparison fixed living environment and more consistent living habit make Effect of Environmental reduce greatly as the material of research multigenic disease.Select such colony,, still therefrom choose family and carry out pedigree analysis, all help to improve the detection sensitivity in hypertension susceptible gene site no matter be to be used for the linkage disequilibrium analysis.Therefore selected in the present embodiment isolated population is as research object (isolated population has the relative uniformity of genetic background, the advantage that founder's number is fewer).

On the basis of informed consent random collecting 324 ages at the Han nationality's individuality more than 50 years old, all come from Dabie Mountain, Yuexi County, Anhui Province and ring the intestines town.80% individuality has only 6 surnames, and founder's number should be fewer, and other individuality of identity comes from same founder.The hypertension sample of selecting for use requires systolic pressure to be not less than 140mmHg, and diastolic pressure is not less than 90mmHg, measures blood pressure and averages for twice.

Select 11% being positioned at the top that blood pressure distributes (37 examples are individual wherein; SBP＞178mmHg) and 7% (22 individualities that are positioned at the lowermost end that blood pressure distributes are arranged; SBP＜104mmHg) is totally 59 individualities, carries out the detection of SNP by the method for direct order-checking.

1.2 experimental technique and result

1.2.1 DNA extraction

Extract DNA with conventional phenol chloroform method from people's blood, concentration correction is used for conventional pcr amplification to 20ng/ul.

1.2.2 the design of PCR and sequencing primer

According to the genome sequence of CAT among the GenBank, design and synthetic following primer.Concrete primer is as shown in table 1 below.

Table 1 primer sequence table

The primer title	Sequence (5 '-3 ')	SEQ?ID?NO：
			Adopted primer is arranged	cttgggtcgaaggctatctg	????2
Antisense primer	ttccacgttcctgtttgtca	????3

1.2.3 the pcr amplification of CAT gene

DNA with extraction is a template, uses the Taq enzyme, carries out pcr amplification with the Touchdown program on GeneAmp 9700 PCR instrument.Reaction conditions is: 94 ℃ of pre-sex change 2 minutes, and 94 ℃ of sex change 30 seconds, 63 ℃ of annealing 40 seconds, 72 ℃ were extended totally 10 circulations, 0.5 ℃ of each cycle annealing lapse of temperature 40 seconds; Later 94 ℃ of sex change 30 seconds were annealed 40 seconds for 58 ℃, and 72 ℃ were extended totally 30 circulations 40 seconds; Last 72 ℃ were extended 7 minutes.Pcr amplification product is verified through agarose gel electrophoresis.

As a result, obtain the amplified production of 402bp.

1.2.4 the discovery of SNP and detection

The PCR product is used ABI-PRISM behind the Resin resin purification ^TM377 dna sequencing instrument (appliedbiosystems of u.s.a. applied biosystem company (ABI)) carry out the two-way order-checking of the terminal cessation method of fluorescent mark, and the interpretation and the SNP that carry out sequence with Polyphred software (the http://droog.mbt.washington.edu/Polyphred.html of Washington, DC university) confirm.

As a result, find to exist following SNP:18934 position T → C.

1.2.5 SNP gene type and association analysis

Carry out the SNP gene type with direct unidirectional sequencing.Promptly in hypertensive patient and normal arterial pressure control group, carry out somatotype and association analysis.

Carry out chi square test after allelic frequency statistics is come out,, determine whether that tentatively allelic frequency and blood pressure level are chain by blood pressure the highest 11% and 7% minimum individual data items are compared.

The frequency of C type SNP in the hypertension individuality that found that among the SEQ ID NO:1 18934 is 32.8%, and the frequency in the ypotension crowd is 16.7%.Difference on the frequency between the two is 16.1%, has confirmed among the SEQ IDNO:1 18934 T/C type SNP and the hypertensive dependency that exists.

Embodiment 2

Essential hypertension susceptibility detection kit

As described in embodiment 1, sudden change and the high blood pressure disease of 18934 T → C are closely related among the SEQ ID NO:1.Therefore, can be that template increases and detects at DNA based on this sudden change design CAT gene-specific primer with patient.

Prepare a test kit (100 person-times), it contains:

Title	Sequence	Concentration
			Adopted primer is arranged	SEQ?ID?NO：2	100pmol
Antisense primer	SEQ?ID?NO：3	100pmol
			The PCR reaction solution	Contain Taq enzyme dNTP magnesium ion PCR reaction buffer

Extract male sex's patients'blood 3ml to be detected, use ordinary method (or using specific test kit) from blood, to extract DNA.PCR primer in the hypertension detection kit is diluted to 1 ì mol/ ì l, is that template is carried out the PCR reaction with the primer that is provided with the DNA that is extracted.Behind the PCR product purification, use ABI-PRISM ^TM377 dna sequencing instrument carry out the two-way order-checking of the terminal cessation method of fluorescent mark, and the interpretation and the SNP that carry out sequence with Polyphred software confirm.

Perhaps, amplified production and normal control are carried out stratographic analysis with sex change high performance liquid chromatograph (DHPLC), also can detect 18934 T → C.

Detected result, the hypertension susceptibility that contains the detected object of 18934 T → C is higher than normal population.

All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.

Sequence table

＜110〉Fudan University

Research Center of Shanghai Human Genome

＜120〉dependency of catalase gene and essential hypertension

<130>040164

<160>3

<170>PatentIn?version?3.1

<210>1

<211>38100

<212>DNA

＜213〉homo sapiens (Homo sapiens)

<400>1

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?ttattagaga?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

tccatttaaa?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?agaggaggag?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?acagattatt????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?attactctaa?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?acacagatct?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?accccatctc?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?gtagttattc?agaagccaat?atttcttaag?caagacatat????7680

taatagtgat?attttgctag?attattgggt?tgacctctaa?aataaatatc?cataaattaa????7740

aatgaaaaat?aggagcttag?tatcagatat?gtgaatccga?gcagcctctg?ccccattctg????7800

aaatcaaaag?gttgtagcta?tgattttatt?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?tttttttttt?tttttttttg?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?ctgggattac????8940

aggcgtgagc?caccgcgccc?agcccatcta?ttttctcaaa?cctattaatc?agtcatttta????9000

aaacctgtat?ctgacagttc?cattatttaa?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

tggtttatat?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?taactttatt???10320

attttattgt?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?ctccacccag?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?acccagatgc?acttaggaat?aagaaaagtc??11400

aagtaagtct?gcaaataatc?actgtgatga?gaaataaatt?caagtggtaa?tcaaatacac??11460

acgtacacat?acacagatat?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?acacatttca?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?caccatacta?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?ttattcactt??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?gtccattttg?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?ggagtttgag?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?cctatactca?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

ttgatttata?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?ggtttactca?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?gtgaagtggt??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?aatagggctt??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?gcggtaggag?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?ttcattatat??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

ccacacttgg?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

ttattatttc?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?ccaccctaac?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?agaggtattt??25680

tgagtctcta?tttagaagtt?tgatgatgtt?ttatcaatca?gaaatatgcc?ataggaactt??25740

aactcttgct?tacgtcaatt?agcctatcat?aaaattggtt?tcattacatg?tcgtttaaag??25800

tcaaagtttc?caagaaccta?ttgatgttaa?gtgaggactt?attgtatttt?tattccttca??25860

atgatgggag?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

ttattactgc?cacgatcttc?aagtgcgcta?ccttgtttag?tgcatctcaa?gaaatgtaca??26760

ccagctattt?gcaacatctc?ttttcaatta?aaaaaaaaaa?gtcaatggct?ttcactgtca??26820

ctaaaagtaa?actgttactg?gtaaggtgga?ttatagtatt?aatgttttaa?agatagcaaa??26880

tgtctctgaa?atgggatagg?ttattttact?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

taggattgaa?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?aaggggggga?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

tatttaagat?gaaaataatt?aatgcaatga?agatttacta?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?ctccatttat?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?cattatcata??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?tcccatttac?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?attataggtg??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?tcccatacac??33480

tgctgatgaa?gtataaaatg?gtacaaccac?tttggaaaaa?ggctcagcag?tttctcataa??33540

cacattcact?taccttatga?cccaataaat?ctactcctag?gtatttccta?agagaaataa??33600

aaccacctgt?gcacaaaaag?acttatgtaa?gaatttttat?agcagcttta?tttattttta??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?agacacaggg??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?tggacaaagg?ggatggctgt??34440

atttcaatta?agagcctccc?aactatagat?gtgttttgga?gtcagccatt?tctcacatgg??34500

gcaactatgt?aaggtaaatc?aacagcaggg?acttcaacag?ttcatgattc?tcccaaacaa??34560

ggaagaaggc?agctagtctc?agaataaagg?ggagtccttt?aaaaatgaaa?atattacacc??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?acccactggg?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

cttgggtcga?aggctatctg???????????????????????????????????????????????????20

<210>3

<211>20

<212>DNA

＜213〉artificial sequence

<220>

<221>misc_feature

＜223〉primer

<400>3

ttccacgttc?ctgtttgtca??????????????????????????????????????????????????20

Claims (10)

1. whether a vitro detection sample exists the method for the single nucleotide polymorphism of catalase gene CAT, it is characterized in that, comprises step:

(a) with the CAT gene of CAT gene-specific primer amplification sample, obtain amplified production; With

(b) detect whether there is following single nucleotide polymorphism in the amplified production:

18934 T → C;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.

2. the method for claim 1 is characterized in that, described gene-specific primer has the sequence of SEQID NO:2 and 3.

3. the method for claim 1 is characterized in that, the length of described amplified production is 100-3000bp, and contains among the SEQ ID NO:1 the 18934th.

4. one kind is detected hypertensive test kit, it is characterized in that it comprises the primer of specific amplification CAT gene or transcript, and it is 100-3000bp that described primer amplification goes out length, and contains among the SEQ ID NO:1 the 18934th amplified production.

5. test kit as claimed in claim 4 is characterized in that, it also contains the reagent that is selected from down group:

(a) with SEQ ID NO:1 in the 18934th sudden change bonded probe;

(b) the 18934th sudden change restriction enzyme among the identification SEQ ID NO:1.

6. test kit as claimed in claim 5 is characterized in that, described sudden change is following single nucleotide polymorphism:

18934 T → C;

Wherein, the nucleotide position numbering is 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. method that the hypertension susceptibility of individuality is diagnosed is characterized in that it comprises step:

Detect this individual CAT gene, transcript and/or albumen, and compare with normal CAT gene, transcript and/or albumen,

There are differences and just show that this individuality suffers from hypertensive possibility and be higher than normal population.

9. method as claimed in claim 8 is characterized in that, detection be gene or the transcript of CAT, and with normal CAT nucleotide sequence comparing difference.

10. method as claimed in claim 9 is characterized in that, described difference is following single nucleotide polymorphism:

18934 T → C;

Wherein, the nucleotide position numbering is based on SEQ ID NO:1.