CN103045625A - Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection - Google Patents
Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection Download PDFInfo
- Publication number
- CN103045625A CN103045625A CN201210531202XA CN201210531202A CN103045625A CN 103045625 A CN103045625 A CN 103045625A CN 201210531202X A CN201210531202X A CN 201210531202XA CN 201210531202 A CN201210531202 A CN 201210531202A CN 103045625 A CN103045625 A CN 103045625A
- Authority
- CN
- China
- Prior art keywords
- myocardial infarction
- reagent
- gene
- vitro detection
- genes involved
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a myocardial infarction relevant gene and a reagent, a preparation or a kit and an application for in-vitro detection, which belongs to the relevant technical field of a myocardial infarction relevant gene in biological engineering. The myocardial infarction relevant gene is a CYP8A1 gene of the eighth mutation; the nucleotide sequence of the gene has a sequence in formula SEQIDNO:1. The invention makes detailed introductions on the myocardial infarction relevant gene and the reagent, the preparation or the kit and the application for in-vitro detection; the kit for the in-vitro detection on the myocardial infarction relevant gene can be used for detecting the polymorphism of the myocardial infarction relevant gene in vitro and also used for detecting, preventing, diagnosing or treating myocardial infarction, so that the happening rate of the myocardial infarction is reduced, thereby being significant in preventing and treating myocardial infarction diseases.
Description
Technical field
The invention belongs to biotechnology Myocardial infarct-related gene-correlation technical field, relate to reagent, preparation or test kit, the application of a kind of myocardial infarction genes involved and vitro detection thereof.
Background technology
Epidemiology and clinical study show, coronary atherosclerotic heart disease (coronary artery disease) is most developed countries and the adult of many developing countries morbidity and main causes of death.Along with the prolongation of the average life span, the affluence of material conditions and the westernization of culture life mode, the trend that increases of the cardiovascular disorder of many developing countries (comprising China), particularly myocardial infarction morbidity and mortality ratio has caused global concern.Expect the year two thousand twenty, myocardial infarction and cerebral apoplexy will become respectively the whole world be ranked first position and the 4th disease.Although the in recent years diagnosis of myocardial infarction and treatment had very great development, thrombolytic drug and interventional therapy etc. has improved the prognosis of myocardial infarction patient greatly, and its nosetiology and pathogenetic research are still made slow progress.Since the sixties in 20th century Frmainghma result of study announce first since, people recognize that progressively myocardial infarction is determined by Other Risk Factors.Except traditional myocardial infarction Hazard Factor, beyond smoking, obesity, hypertension etc., the interaction of inherited genetic factors and environmental factors plays very important effect in the genesis of myocardial infarction.
Myocardial infarction (myocardial infarction; MI) be a kind of serious state of coronary heart disease, disability rate and mortality ratio all occupy first of the coronary heart disease.Along with the development of Protocols in Molecular Biology, the effect of inherited genetic factors in the myocardial infarction pathogenesis becomes the focus of present research gradually.In recent years fundamental research shows, the variation of some gene comprises that renin-angiotensin-aldosterone system, lipid metabolism system, blood coagulation and the gene pleiomorphisms such as fibrinolytic system and inflammatory factor can increase the occurrence risk of coronary heart disease.
Prostacyclin (prostacyclin, PGI2) at first be found in 1976, it is arachidonic a kind of meta-bolites, in normal ph water, be hydrolyzed into the 6-ketone-PGF1α (6-ketone-PGF1 α) of non-activity, transformation period is about 6 min in human body, main generation by stimulating ring adenosine phosphate (cAMP) causes to relax the VSM and suppresses the growth of smooth muscle cell, powerful antiplatelet aggregative activity is arranged simultaneously.In addition, PGI2 can also impel endotheliocyte to discharge nitrogen protoxide (NO), and NO can impel PGI2 to generate increase, and as seen both can form benign cycle, the common diastole that participates in keeping vascular smooth muscle.Therefore just consider its application problem in cardiovascular field from PGI2 people after being found in 1976; there was the investigator to propose as far back as 1979; PGI2 has the cytoprotections such as the myocardial infarction area of minimizing and myocardial consumption of oxygen, but mechanism is not also illustrated fully.At present sure be PGI2 to the therapeutic action of pulmonary hypertension, and there have PGI2 preparation and its analogue to be applied to be clinical, and obtain sure curative effect.The effectively generation of preventing cardiovascular disease of PGI2 analogue is thought in Japanese scholars Nakayama research.Because the effect of its vasodilator, anticoagulant and anti-smooth muscle cell proliferation, thereby block some link of atherosclerosis genesis, be conducive to the generation of prevention of arterial thrombotic diseases.Therefore it is contemplated that the decline of PGI2 level can reduce it increases arterial thrombotic disease to the provide protection of blood vessel incidence.
Prostacyclin synthase (prostacyclin synthase, PGIS) is the member in the Cytochrome P450 superfamily, can generate PGI2 by catalysis PGH2 (PGH2).Studies confirm that PGIS mainly is distributed in vascular endothelial cell and smooth muscle cell, but studies show that recently PGIS also has extensive distribution in central nervous system, Purkinje cell such as pallium and pyramidal cell of the hippocampus and cerebellum, the author finds that also PGIS mRNA all has great expression in blood vessel and maincenter large neuron, and the two has obvious dependency.Other has experimentation on animals proof PGIS cDNA transfection to rat smooth muscle cells can cause the synthetic growth that increases and suppress cell of PGI2.The results of study such as Zou show that also the PGIS activity decreased can promote atherosclerotic generation.The above results shows that PGIS not only can regulate the synthetic of PGI2, but also has the effect of direct inhibition vascular smooth muscle cell proliferation, and may be significant to the adjusting of central nervous system function.The generation of PGI2 mainly is subject to the adjusting of PGIS, and the CYP8A1 gene is the gene of coding PGIS, and its genetic polymorphism significantly affects the synthetic existence of PGI2.The PGIS assignment of genes gene mapping is on No. 20 karyomit(e), and length is approximately 60 kb and comprises 10 exons.Japanese scholars Nakayama etc. carried out order-checking and its structure was carried out system description the PGIS gene the same year.Think that the length of PGIS gene is 70kb, be about 30 times of cDNA length, the length range of exon is 74bp to 182bp, and intron is 1kb to 19kb.
The a large amount of effect of clinical research confirmation PGI2 in cardiovascular disorder arranged in recent years, and the height of its serum level may determine directly that the risk of myocardial infarction occurs the patient.This protein might be by the prediction index of priority application in the ill risk of myocardial infarction clinically.
Summary of the invention
The invention provides reagent, preparation or test kit, the application of a kind of myocardial infarction genes involved and vitro detection thereof, overcome the deficiency of above-mentioned prior art, it can effectively solve prior art undesirable problem of effect when prevention and treatment myocardial infarction.
One of technical scheme of the present invention realizes by following measures: a kind of myocardial infarction genes involved i.e. the CYP8A1 gene of the 8th exons mutation, and the nucleotide sequence of this gene has the sequence shown in the SEQ ID NO:1.
Two of technical scheme of the present invention realizes by following measures: the reagent of a kind of vitro detection myocardial infarction genes involved according to claim 1, this reagent is for detection of the polymorphism in CYP8A1 gene G1120C site, and this reagent comprises following primer:
Upstream primer: 5'ATGTGACTGTGTGGGCGAG3';
Downstream primer: 5'AGGAAGGGGAAGAGGAGGA3'.
The below is two further optimization and/or improvements to the foregoing invention technical scheme:
Mentioned reagent can be the reagent that combines with the restriction fragment length polymorphism analysis for the polymerase chain reaction or is used for the reagent that the polymerase chain reaction combines with direct sequencing.
Three of technical scheme of the present invention realizes by following measures: a kind of preparation or test kit that the reagent of vitro detection myocardial infarction genes involved is arranged, this test kit comprises pcr amplification enzyme and corresponding damping fluid.
Four of technical scheme of the present invention realizes by following measures: a kind of reagent of vitro detection myocardial infarction genes involved is for the preparation of the preparation of vitro detection myocardial infarction genes involved or the application in the test kit.
The below is two further optimization and/or improvements to the foregoing invention technical scheme:
Can comprise pcr amplification enzyme and corresponding damping fluid in the mentioned reagent box.
The present invention is to reagent, preparation or the test kit of myocardial infarction genes involved and vitro detection thereof and should be used as detailed introduction, the test kit of this vitro detection myocardial infarction genes involved is except being used for the polymorphism of vitro detection myocardial infarction genes involved, can also for detection of, the prevention, the diagnosis or the treatment myocardial infarction, thereby reduce the incidence of myocardial infarction, prevention and treatment myocardial infarction disease are significant.
Description of drawings
Fig. 1 is the genome structure schematic diagram of Myocardial infarct-related gene of the present invention, and as can be seen from the figure the myocardial infarction genes involved is made of 10 exons and 9 introns.
Fig. 2 is the examination schematic flow sheet of Myocardial infarct-related gene order variation of the present invention.
Fig. 3 is part order-checking collection of illustrative plates of the present invention.
Fig. 4 is the restriction enzyme mapping of G1120C pleomorphism site of the present invention.Wherein show that such as swimming lane 3 genotype in individual G1120C to be measured site is the C/G heterozygote; Swimming lane 1 shows that the G1120C site of individuality to be measured is the C/C homozygote; Swimming lane 2 shows that the G1120C site of individuality to be measured is the G/G homozygote.
Embodiment
In order more clearly to understand the present invention, further describe the present invention referring now to the following example and accompanying drawing, embodiment only is used for explaining and does not limit the present invention in any way.The experimental technique of unreceipted actual conditions among the embodiment, usually carry out according to normal condition: " molecular cloning: laboratory manual " (the New fork:Cold Spring Harbor Laboratory Press that shows such as people such as Sambrook, 1989) condition described in, or the condition of advising according to manufacturer.
Upstream primer: 5'ATGTGACTGTGTGGGCGAG3';
Downstream primer: 5'AGGAAGGGGAAGAGGAGGA3'.
This CYP8A1 gene and registration number in Genbank are that the CYP8A1 gene of 5740 (Gene ID) is compared, and are the 8th exons mutation; Particularly, for the allelotrope of G1120C pleomorphism site is C, C1120C A/G pleomorphism site is according to the definition of the position in the CYP8A1 genome, is to count from the transcription initiation site of CYP8A1 gene to be that the+1,1120th base is C specifically.
So-called " gene pleiomorphism " refers in the crowd, the difference that the nucleotide sequence of each genes of individuals exists.Those of ordinary skills are known, and pleomorphism site of the present invention is single nucleotide polymorphism (SNP) site, and namely mononucleotide changes in the genome sequence; The difference of nucleotide acid sequence can be embodied on the dna level or on the rna level, so myocardial infarction genes involved of the present invention can be embodied in dna level, and on the rna level, preferred dna level, more preferably genomic dna.
The at present research about the complex character disease has two kinds of strategies, the method for linkage analysis and case-control study.Because myocardial infarction is a kind of multi-factor disease, is by the coefficient result of h and E factor, thereby adopt comparatively difficulty of linkage analysis.From the angle analysis of heredity, coronary heart disease/myocardial infarction belongs to complex inheritance proterties disease.In the factor that works to this disease, except the environment factor, the gene number that works is estimated to reach up to a hundred, exists again interaction between each gene, and gene again and exist between the environment and interact.Coronary heart disease/myocardial infarction have certain familial aggregation, but more be some Sporadic cases.What myocardial infarction occured mostly is some Aged Patients, often waits death of patient when knowing, therefore is not easy to collect very much the family data of myocardial infarction.So the present invention adopts case control study.Described case control study is exactly certain allelic frequency of (case and contrast) comparison in two groups of crowds that adopt random choose.
The present invention's statistical study by large sample in the Wei Weier clansman group of Xinjiang (500 routine myocardial infarction patients and 500 example contrasts), through a large amount of experiments, at last proved the CYP8A1 gene with G1120C pleomorphism site of the present invention as the myocardial infarction genes involved take conclusive evidence, the danger that the allelic carrier of C suffers from myocardial infarction has increased by 2 times than G allelotrope carrier.
Embodiment 4, and a kind of preparation or test kit that the reagent of vitro detection myocardial infarction genes involved is arranged is characterized in that this test kit comprises pcr amplification enzyme and corresponding damping fluid.The test kit that the present invention detects the myocardial infarction genes involved of the 8th exon region polymorphism can be used for the polymorphism of vitro detection myocardial infarction genes involved; The test kit of the myocardial infarction genes involved of the 8th exon region sudden change can be for detection of, prevention, diagnosis or treatment myocardial infarction; The method of vitro detection myocardial infarction genes involved can for detection of, the prevention, the diagnosis or the treatment myocardial infarction.
Embodiment 5, and a kind of reagent of vitro detection myocardial infarction genes involved is for the preparation of the preparation of vitro detection myocardial infarction genes involved or the application in the test kit.The test kit that the present invention detects the myocardial infarction genes involved of the 8th exon region polymorphism can be used for the polymorphism of vitro detection myocardial infarction genes involved; The test kit of the myocardial infarction genes involved of the 8th exon region sudden change can be for detection of, prevention, diagnosis or treatment myocardial infarction; The method of vitro detection myocardial infarction genes involved can for detection of, the prevention, the diagnosis or the treatment myocardial infarction.
Embodiment 6, are with the difference of above-described embodiment, and test kit also comprises pcr amplification enzyme and corresponding damping fluid.
Embodiment 7, the choosing of research object
Myocardial infarction group (MI group) 500 examples are selected from January, 2006～2010 year March at No.1 Hospital Attached to Xinjiang Medical Univ.'s Heart center inpatient, meet the World Health Organization (WHO) about name and the Case definition of ischemic heart disease.Control group 500 examples, the examinee of myocardial infarction is got rid of in the underwent coronary contrast examination of selecting to be in hospital the same period.Two groups are the crowd of the Uygur nationality, and age and sex composition have harmony than all mating.
The Case definition of the ischemic heart disease that inclusive criteria: MI group is formulated according to the World Health Organization select the new MI case within the morbidity 3 months, and all the underwent coronary radiography confirms to have one or Multivessel obturation or Serious Stenosis.Control group selects to have coronary angiography data and result to confirm the examinee that is in hospital without coronary artery pathological changes.Two groups of signature Informed Consent Forms before including research in.
Rejecting standard: MI group: the full person of clinical data and the one that merges simultaneously following disease are rejected.(as: dissection of aorta, rheumatic heart disease, Congenital Heart patient, MOFE and have mental disorder can not cooperation person).Control group: spiritedness obstacle person or check through doppler and to confirm that carotid plaques or narrow person are arranged, rejected.
Embodiment 8, the examination of myocardial infarction genes involved sequence variations
Whole myocardial infarction genes involved sequence variations examination flow process is seen Fig. 2.Random choose 48 routine myocardial infarction patients consist of 96 karyomit(e)s as the examination object from the case group, and this sample size can provide 95% power of a test to detect rare gene frequency greater than 1% sequence variations.All individualities are the Uygur nationality, and without any genetic connection.CYP8A1 exon sequence according to announcing among the NCBI relatively obtains the CYP8A1 gene order by the blast database.The method that detects polymorphism is to utilize PCR product direct Sequencing.Utilize Primer 5.0 software design PCR primers, the amplification whole exons of CYP8A1 gene and exon-intron junction region.Primer sequence sees Table 1.
Reaction system (25 μ l): genomic templates DNA 50ng is (from blood preparation, according to ordinary method intraleukocytic DNA is extracted with the imitative method of phenol-atmosphere or with salting-out process and to get final product), 10 * PCR Buffer, 2.5 μ l, dNTPs1 μ l (10 mmol/L), each 0.5 μ l(20 pmol/ μ l of primer P1, P2), dna profiling 1 μ l, Taq archaeal dna polymerase 2.5 U(0.5 μ l).In the reaction of the enterprising performing PCR of 96 hole PCR automatic circulation instruments, PCR loop parameter: 96 ℃ of denaturations 5 minutes; Through 94 ℃ 30 seconds, 58-62 ℃ 45 seconds, 72 ℃ 1 minute; Circulation was extended 10 minutes in 72 ℃ after 35 weeks.The PCR product carries out sequencing reaction behind conventional purifying 2 times, with the PCR primer as sequencing primer.Order-checking is finished at ABI 3500 sequenators.Fig. 3 display part order-checking collection of illustrative plates.
Result: take the Xinjiang, China Uygur nationality as research object, through the sequence screening of system, from the CYP8A1 gene order, find new SNP site, according to current naming rule called after G1120C, be positioned at the 8th exon 1 of CYP8A1 gene order.
Embodiment 9, adopt the PCR-RFLP method to detect myocardial infarction genes involved G1120C polymorphism of the present invention
Method: PCR reaction system (25 μ l): genomic templates DNA 50ng is (from blood preparation, according to ordinary method intraleukocytic DNA is extracted with the imitative method of phenol-atmosphere or with salting-out process and to get final product), 10 * PCR Buffer, 2.5 μ l, dNTPs1 μ l (10 mmol/L), each 0.5 μ l(20 pmol/ μ l of primer P1, P2), dna profiling 1 μ l, Taq archaeal dna polymerase 2.5 U(0.5 μ l).In the reaction of the enterprising performing PCR of 96 hole PCR automatic circulation instruments, PCR loop parameter: 96 ℃ of denaturations 5 minutes; Through 94 ℃ 30 seconds, 58-62 ℃ 45 seconds, 72 ℃ 1 minute; Circulation was extended 10 minutes in 72 ℃ after 35 weeks.Primer is as follows:
5’ATGTGACTGTGTGGGCGAG3’
5’AGGAAGGGGAAGAGGAGGA3’
Amplified production length is 275 bases, selects EcoR I restriction enzyme (Lithuania MBI company) to carry out enzyme and cuts.The enzyme tangent condition is referring to the operational manual of manufacturer.Then observing length with ultraviolet transilluminator behind the electrophoresis on 2% the agarose gel.
The result: as shown in Figure 4, when having produced 230 and during the fragment of 45 two kind of length after the long PCR product cutting of 275 bases, the allelotrope that the G1120C site be described is G, and when the PCR product that only has 275 bases to grow, the allelotrope that the G1120 site is described is C.Because testing sample shown in Figure 4 is the DNA from individuality to be measured, so, shown in swimming lane 3, produce 3 kinds of fragments, then the genotype in this individual G1120C to be measured site is the C/G heterozygote; Carry two kinds of allelotrope of C and G; Shown in swimming lane 1, only have a fragment, the G1120C site of this individuality to be measured is the C/C homozygote, carries C allelotrope; Shown in swimming lane 2, only produce 2 useful fragments, the G1120C site of individuality to be measured is the G/G homozygote, carries G allelotrope.
Embodiment 10, utilize PCR and direct sequencing to detect G1120C polymorphism of the present invention
Method: at first the PCR reaction is carried out in the G1120C site, actual conditions and primer are referring to embodiment 3.The PCR product is directly read sequence on ABI 3500 sequenators.
Result: as shown in Figures 2 and 3, be order-checking process of the present invention and part sequencer map.Wherein 1 to 3 show that respectively the G1120C pleomorphism site of the present invention's individuality to be measured is, CC homozygote, C/G heterozygote and GG homozygote in the sequencer map of Fig. 2.
Embodiment 11, the association study of polymorphic site and myocardial infarction
For further inquiring into relation between c reactive protein gene and the myocardial infarction morbidity from the angle of heredity, and provide the evidence of genetic epidemiology for existing similar research both at home and abroad, use the methods of genotyping of the current international practice of PCR-RFLP, in the 500 routine myocardial infarction patients of collecting and corresponding 500 example contrasts, vitro detection is from the polymorphism in G1120C site in the sample of individuality to be measured, thereby analysis G1120C site is at myocardial infarction patient and normal control crowd's distributional difference.
At first use the Hardy-Weinberg balance check.The Hardy-Weinberg balance is a kind of concept of population genetics: mainly refer to a group human body that a nationality lives in a certain area, can mutually hybridize at random, whole genetic information that this colony has are called the gene pool of this colony, it directly reflects this area, the hereditary feature that this is national.Expression-form gene frequency and the genotype frequency of these genetic information, under the condition that does not have sudden change, migration and genetic drift, gene frequency and genotype frequency are observed to remain unchanged and are the Hardy-Weinberg balance in the colony.Existing like this genetic polymorphism possesses again hereditary stability.
The gene type interpretation of result is found, the gene type result meets the Hardy-Weinberg balance, therefore can get rid of experimental error, and the result is reliable for this gene type.Analytical results sees Table 2.
Conclusion: single factor analysis is found, three kinds of genotype individualities of polymorphic site G1120C are distributed with notable difference (P＜0.05) in case group and control group, the aobvious recessive inheritance pattern of C/G is further analyzed discovery, the allelic carrier of the relative G of the allelic carrier of C has marked difference (P＜0.05) in case group and control group, wherein the allelic carrier of C to suffer from the danger of myocardial infarction be 2 times of the allelic carrier of the G danger of suffering from myocardial infarction.
Disturb the most serious factor of case control study to comprise crowd's mixing in hereditary basis, when case is selected because the disease that standard does not strictly produce mixes etc., more than having got rid of on the basis of several Confounding Factor, the contriver has selected 500 research samples and corresponding contrast, this belongs to larger research sample in the similar research in the world, in order further to analyze the G1120C site to the contribution of myocardial infarction risk level, adjusting the age, biochemical indicator, hypertension, after the impact of other Hazard Factor of myocardial infarction such as diabetes, carry out the multivariate logistic analysis of regression model.The logistic analysis of regression model is a kind of common analytical procedure statistically, and it is mainly used in the analysis of dichotomic variable, can be used for adjusting whole analysis is caused the factor that mixes.The results are shown in Table 3.
The result: as shown in table 3, multivariate logistic s analysis of regression model shows that G1120C still remains the Hazard Factor of myocardial infarction.
Conclusion: the CYP8A1 gene that the sudden change of the 8th exon region is described is the myocardial infarction genes involved, and for c reactive protein G1120C site, carry the individual danger of suffering from myocardial infarction of C allelotrope and significantly raise than carrying allelic individual the dangerous of myocardial infarction of suffering from of G.
Embodiment 12, and polymorphic site G1120C is the evidence on the function of an inheritance susceptible factor of myocardial infarction
G1120C itself is positioned at the CYP8A1 gene coding region, and the sudden change in this site belongs to a kind of nonsynonymous mutation, probably can affect level or the activity of PGIS.And then affect its downstream product PGI2 concentration, thereby the genetic predisposition that different individualities is produced myocardial infarction increases.For the specific target site of this site design medicine can establishment because the level of the PGI2 that transgenation causes descends, suppress vasoconstriction and platelet aggregation, thereby reduce the incidence of myocardial infarction.This will produce huge economic benefit.
The contriver has studied the level of PGI2 between the different genotype, found that, C allelotrope carrier and G allelotrope carrier relatively, blood plasma 6-ketone-PGF1 alpha levels obviously reduce (
P＜0.05).
Conclusion: the CYP8A1 gene that further specifies the 8th exons mutation is the myocardial infarction genes involved.This site is found the new drug development tool is of great significance.
Embodiment 13, and vitro detection myocardial infarction genes involved is the test kit of CYP8A1 gene
A kind of test kit of vitro detection myocardial infarction genes involved, this test kit comprises:
1) primer in amplification G1120C site:
5’ATGTGACTGTGTGGGCGAG3’
5’AGGAAGGGGAAGAGGAGGA3’
2) pcr amplification enzyme and corresponding damping fluid;
3) EcoR I restriction enzyme and corresponding restriction enzyme mapping.
The gene order table:
Application Project
＜120〉Title: the reagent of myocardial infarction genes involved and vitro detection thereof, preparation or test kit, application
<212> Type : DNA
<211> Length : 61892
SequenceName: the myocardial infarction genes involved i.e. the CYP8A1 gene of the 8th exons mutation
<213> OrganismName : CYP8A1
<400> PreSequenceString :
actcacttat ctacctatcc attcatctat ccatccgccc atccattcat ccatccaccc 56340
accaatttac cctttcatcc acccatctac ccattcattt atccatccag ccacccatcc 56400
actcatctat ccatccatcc actcatctac ccatgcatcc atctgtccat tcatccattc 56460
acaaattcac ccatccatcc attaatctat ccatccaccc atgcatccat gcatccatcc 56520
atccatccac atactcaccc atctacccat ccattaatct atccatccac ccatccatcc 56580
atccatccac ccatgaagct actctttcat ccacccacct acccatgaat ccatccatct 56640
atccatccgt ctgtccattc aatcatctac tcatttatcc atccatccac ccactcattt 56700
atctacccac tcactcatat gtccatccac ccattcttct acttatccac ccatccatcc 56760
atctatccac ccatctactc atccatccat ccatttgtcc attcatccat ttattcatcc 56820
atccacccat caatctaccc attcattcac ccatctaccc atccatccat ccaccccatc 56880
tacccatcca tccatctatc catccctcca ttcatccatc tttccatcca catttccatc 56940
tcttagtcta tccactggtt cattcaccta tccttccacc catctttcag tcggcagcta 57000
ctccatgcca agctctgtgc tcagctacaa cccttgcctt accactgtgc agtgtaacca 57060
aaggaaattg gcattttctt tttaaaaaat attttaattt cttttatttt acattacaaa 57120
agtaatatat atcatagaat ataaagaaat acagagaaat atatacttaa atgtaaaaaa 57180
tatgtatttc ccttcttctc cagttctaca gagttctctg accaaggttt ggagtgtgcc 57240
ttgcgaattg ttttcatggc tttacaggca tacacacaac acattcacat gccatgaaaa 57300
gattggtgtg tgtataggat catgcaatat atactattct gaaacttgct tttttcactc 57360
aacaaccaat tgtagaaatt ctttccatgt cagtacacac aaaccttttg ccataatttt 57420
ttaaccaatc ccctatcaat tgacctgtat atggtttcca gtttttccct cttataaaca 57480
tgactgtcat gaacatcctc atacattttt cttggtgtga gcttgttctg ggaatggggt 57540
cagaaaagga ccttcttcct atttcaggta tttaaataca accgattcct gaaccctgay 57600
ggatcagaga agaaagactt ttacaaggat gggaaacggc tgaagaatta caacatgccc 57660
tggggggcgg ggcacaatca ctgcctgggg aggagttatg cggtcaacag catcaaacag 57720
tgagtggggg gccagggcag ggccagggtg gctcctgatc ccctggttgg caggcgccct 57780
tctctcctgg gactgagggc tcagccagga gaatggtttg gcagaggctt cacatggaca 57840
gcccgtgagc catctcttat ttgtggtttt acaactgtgt gcttttataa atcagaagag 57900
ctcgtgtaaa aaagccagat ttcggatttc ttttgaaata atcagatctg gcagcaggtt 57960
cttccctcca agactggtgt ccagattcca aatactggac attcctttca gctgaccgta 58020
gctacccgtc tgcagctgct gccgcatgtt cagaacttca tgttaggaac tggccgaact 58080
tcactgttcc cacttggtta ttttaatacc tgtggggccg agagcctcat aaattatctc 58140
ccttgaaccc caggagcttt ggccaccagg atcacaaaca gattggacca ggggttatga 58200
catctgccct gtcctcctgt ccagcccctg tgcaatcata agctgggtta ccttctgagc 58260
ctgtttcctc aactgtaaaa tgagggttct gattataact ataccccagg agtgtcagga 58320
taaatgaatt ggatctttct aagttcttaa cccactgtct ggctcattgt aggcatttga 58380
aaaatgttaa ttattaccat aattgttaat atcattattg tgcatgcatt gtgctaagtg 58440
agtgggcacg ttacataaac ctttagcctc agtttcctca agaaaatgga gtcaattatg 58500
aacataatac agctcatatg cattccttaa aatcaccttg ctgtgtgaaa tcatgcaaca 58560
aaaactgcag ggattcacgc ctgtaatccc agcactttgg gaggccgagg tgggcagatc 58620
atttgaggcc aggagttgga gaccagccta gccaacatgg tgaaaccccg tctctactaa 58680
aaatacaaaa attggccggg tgtggtggag cgtgcctgta atcccaacta ctcaggaggc 58740
tgaggcagga gaatctcttg aacccaggag gtggagattg cagtgagcca agattgcgcc 58800
gttgcactcc agcctgggcg acagagtgag actctgtctc cagaaaaaaa aaaaaaaaaa 58860
aaaaagaaag aaagaaagaa aaaactgcag ggattgaggg aaaaggggga ctggggccaa 58920
cactcaaaaa cctcatcagt gacataataa aaaaaaaaga gagagagaga agaacccagt 58980
aaaaaaaatg gtagtaaagt tttatacatg ttacctggct aagaactatg taagtgctgt 59040
aataactgtg gctgtagatc gatgcttagt tgtgtgcatg tgtatatttt atggatttct 59100
gcacacccag gttcagctgg gtgcagtctt tcgtcttcac ctcatgtttc tcacaagtga 59160
aattgcacgt gagcaaatgt gaaacttgca ttatattcaa atggtgccct aatatatcaa 59220
tcctattgga acaaattcac ctgttcaaaa caagtgttag agcagaaccg actggcattc 59280
ctacctcatc aagtttttgg acagataaaa agagatagcg ggtactaaat gcttagatgg 59340
tgcctagcgc acagtgttcc aataaattag ctaatttgtt gttattttta tctcactgaa 59400
ttgtcatgac aaccctgctg gtccatctaa aaccgagtct aggcaggtca gatcatttgc 59460
cgtgttccca ggaagtggag gtgcaggagt tcaaatccag gggtgggcac aaaaccgcca 59520
tcccttagaa actcaccctc tagtttccta cagcctcaga gctggaagag actcaacatg 59580
cctttcccct ctttgaccag atgggagaac tgagacccag agtgggtaag aacttggcga 59640
gggccacatt cccataagcg gcacagccca gatctgtcca gagtgttaac tcccctctct 59700
cctcatctgc cctcttcttc actccttcag tgagacaaaa gaatccccac catgtgctca 59760
gtgctgcatc gggcgcctgg gttggggtag ggaatagtgg tgggaaaaga gtcataagat 59820
attctcctga cccttacgaa catgggcatt ggtaagatga ggcagggagg cagcaaccgt 59880
tgacatcccc cgtagggctg cctccacatt tatgggggca ctgattacat cataggcatg 59940
gcagctgcga atagttatta tgacagtttt cccagaagat ggcagtagag agtgcttgtt 60000
cttgcaaagt tggtctattt aggacaattt tctcagagat ggcagtaaaa tgtcttgaga 60060
gcgatgcctg tttctaatct cacaaagaca cctaagggtt agcagcagtc ctggccaaga 60120
tagacagtga gggttccagg agtgtgggga aaggcggcaa agtgtgtggg ctggaggatc 60180
cagaggagtc gccgtattaa tcagcacagg ctcagctctg cacctgccga gtctcagtgg 60240
cttttaccaa cagaggtgta gttccactca cacggtatca gctgtgggat tggagggctt 60300
ccccactctg tgcacagact cgggccaggc tctttccctc tttcggctcc accgtctcaa 60360
tgtgaagcct gcatggctgc caggcaggag ggacagctgg agagccacac actggctcgc 60420
gaatgctttg acacaggagt gacacaggcc acttctgcac acagcccatt caccagaact 60480
catccacggg cctgcctaac cttacagggg gctggcaaat gtggtgagca gcaaatgcgt 60540
ttcttctact tcctcactcc tctcctcact ttactcaaag ttggagtcag acttaggagg 60600
cagccatggc tcccctgcct tgttcccgat aaccccctgc cctactcctc tccttcacac 60660
cttccatgcc atctctctgc agatttgtgt tccttgtgct ggtgcacttg gacttggagc 60720
tgatcaacgc agatgtggag atccctgagt ttgacctcag caggtacggc ttcggtctga 60780
tgcagccrga acacgacgtg cccgtccgct accgcatccg cccatgacac agggagcaga 60840
tggatccacg tgctcgcctc tgcccagcct gccccagcct gccccagcct cccagctttc 60900
tgtgtgcaca gttggcccgg gtgcaggtgc tagcattacc acttccctgc ttttctccca 60960
gaaggctggg tccaggggag ggaaaagcta agagggtgaa caaagaaaag acattgaaag 61020
ctctatggat tatccactgc aaagttttct ttccaaaatc aggctttgtc tgctcccaat 61080
tcacctcgtt actctcacct cgtgatatcc acaaatgcta ttcagataag gcagaactag 61140
gagtcttcac tgctctgccc ccaactcccg gaggtgtcac cttcctagtt cttatgagct 61200
agcatggccc gggccttatc cagtcaaagc ggatgctggc cacagaaagg ccactcagga 61260
tgtcctttgt gtccattgat gtcattcagc agtcagtccc ccaataatcc ttaaactagc 61320
taaaaccaaa ggagtccctt agaagatctg cttccctggg gccccatttg ccagattgcc 61380
ccattgctca cactacttga gaaaatgcag gagagcttcc cccaaggctg atgcattccc 61440
ggtgcagaac aggggcaccc tccaaacact gggctctgag gagtggagtt ctctgttcta 61500
gagtgacagg caccagatgg gatgggcttt ctcagtgtca gcactcaggt agggagctaa 61560
ggaagacaca gcccagacaa gatggctgga aggagccagc caggactcct tagactgatc 61620
aagccaaaaa agaaggtgcc gatttcatgc attctagtgc agaagcccca actgtgatca 61680
cgatccagtc tgcagacgtg ttttgtttgg acttcactta aaaaaatgcc ttagttgtta 61740
tcatctttgg gagagttcat tcaaaatgtc cagcttctct tgaaaacttg gtatatctgg 61800
ccacactggg ctcacattcc caagggtaac tcttggccag agctgagtgg cagccgcctc 61860
ccttatgcag gacatgtgct ctcggcttca cc 61892
Claims (6)
1. the CYP8A1 gene that the myocardial infarction genes involved is the 8th exons mutation is characterized in that the nucleotide sequence of this gene has the sequence shown in the SEQ ID NO:1.
2. the reagent of a vitro detection myocardial infarction genes involved according to claim 1 is characterized in that this reagent for detection of the polymorphism in CYP8A1 gene G1120C site, and this reagent comprises following primer:
Upstream primer: 5'ATGTGACTGTGTGGGCGAG3';
Downstream primer: 5'AGGAAGGGGAAGAGGAGGA3'.
3. the reagent of vitro detection myocardial infarction genes involved according to claim 2 is characterized in that this reagent is the reagent that combines with the restriction fragment length polymorphism analysis for the polymerase chain reaction or the reagent that combines with direct sequencing for the polymerase chain reaction.
4. preparation or the test kit of the reagent of a with good grounds claim 2 or 3 described vitro detection myocardial infarction genes involveds is characterized in that this test kit comprises pcr amplification enzyme and corresponding damping fluid.
One kind according to claim 2 or the reagent of 3 described vitro detection myocardial infarction genes involveds for the preparation of the preparation of vitro detection myocardial infarction genes involved or the application in the test kit.
6. application according to claim 5 is characterized in that comprising in the test kit pcr amplification enzyme and corresponding damping fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210531202XA CN103045625A (en) | 2012-12-11 | 2012-12-11 | Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210531202XA CN103045625A (en) | 2012-12-11 | 2012-12-11 | Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103045625A true CN103045625A (en) | 2013-04-17 |
Family
ID=48058486
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210531202XA Pending CN103045625A (en) | 2012-12-11 | 2012-12-11 | Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103045625A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112964876A (en) * | 2021-01-29 | 2021-06-15 | 泰达国际心血管病医院 | New application of prostaglandin H2D-isomerase antibody and ST-elevation type myocardial infarction early screening and diagnosing kit |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101947223A (en) * | 2010-08-25 | 2011-01-19 | 新疆医科大学第一附属医院 | Composite anesthetic, preparation method thereof, and use thereof in myocardial infarction animal model |
-
2012
- 2012-12-11 CN CN201210531202XA patent/CN103045625A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101947223A (en) * | 2010-08-25 | 2011-01-19 | 新疆医科大学第一附属医院 | Composite anesthetic, preparation method thereof, and use thereof in myocardial infarction animal model |
Non-Patent Citations (4)
| Title |
|---|
| NAKAYAMA T ET AL: "Association of a novel single nuclcotide polymorphism of the prostacyclin synthase gent with myocardial infarction", 《AM HEART J》 * |
| 李靖等: "冠心病新致病基因MEF2A第一、第八外显子突变的检测", 《中国优生与遗传杂志》 * |
| 谢翔: "前列环素合酶、环氧化酶-2基因多态性与疆维吾尔族人群心肌梗死的关联研究", 《中国优秀硕士论文全文数据库》 * |
| 谢翔等: "环氧化酶-2基因-765G>C和前列环素合酶基因C1117A多态性与新疆维吾尔族人群心肌梗死的相关性研究", 《中国流行病学杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112964876A (en) * | 2021-01-29 | 2021-06-15 | 泰达国际心血管病医院 | New application of prostaglandin H2D-isomerase antibody and ST-elevation type myocardial infarction early screening and diagnosing kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jiang et al. | A polymorphism in the promoter region of catalase is associated with blood pressure levels | |
| Aoki-Suzuki et al. | A family-based association study and gene expression analyses of netrin-G1 and-G2 genes in schizophrenia | |
| JP4516990B2 (en) | How to predict the range of warfarin dosage | |
| Barton et al. | PLAUR polymorphisms are associated with asthma, PLAUR levels, and lung function decline | |
| Gao et al. | A polymorphism of matrix Gla protein gene is associated with kidney stones | |
| Greenwood et al. | Segmental linkage disequilibrium within the dopamine transporter gene | |
| Xie et al. | Haplotype analysis of the CYP8A1 gene associated with myocardial infarction | |
| JP2009511026A (en) | Method for diagnosing thromboembolic and coronary heart disease | |
| JP4496167B2 (en) | Predisposition detection method for high altitude pulmonary edema | |
| CN101245392B (en) | Method and reagent kit for forecasting susceptibility of incidence of intracerebral haemorrhage | |
| CN103667301A (en) | Gene related to coronary heart disease, and in-vitro detection reagent, preparation or kit and application thereof | |
| Jia et al. | Joint effects of eNOS gene T-786C and ADH2 Arg47His polymorphisms on the risk of premature coronary artery disease | |
| CN103045625A (en) | Myocardial infarction relevant gene and reagent, preparation or kit and application for in-vitro detection | |
| CN104513828B (en) | Coronary heart disease dependent genes are reagent, preparation or the kit of CYP17A1 genes and its vitro detection, application | |
| CN100545170C (en) | A kind of coronary heart disease dependent genes and detection method thereof and purposes | |
| Chang et al. | Association of TSC gene variants and hypertension in Mongolian and Han populations | |
| CN1891823B (en) | CETP gene with specific mononucleotide pleiomorphism, and its detecting method and use | |
| Okada et al. | Identification of protein Sα gene mutations including four novel mutations in eight unrelated patients with protein S deficiency | |
| JP2005511095A (en) | Bi-allelic marker of D-amino acid oxidase and use thereof | |
| Wei et al. | Searching for a locus for schizophrenia within chromosome Xp11 | |
| CN104372004B (en) | One detection method and its application with the susceptible relevant miR-27a Polymorphisms site of myocardial infarction | |
| JP5489146B2 (en) | Genetic risk detection method for obesity | |
| KR20060096437A (en) | Genetic markers for obesity | |
| CN103173454B (en) | Polymorphic sites of obsessive-compulsive disorder related gene GalR1 (Galanin Receptor 1) and application thereof | |
| JP5002746B2 (en) | Gene polymorphisms useful for predicting responsiveness to antidepressants |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130417 |