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Objective To investigate the association between the three single nucleotide polymorphisms (SNP) in angiotensin Ⅱ type one receptor (AT1R) gene with coronary artery disease (CAD) in Chinese Han nationality.Methods Extracted DNA and RNA samples of peripheral blood white cells from 192 CAD patients and 189 healthy individuals in Jan 2011 to Oct 2013 from the general hospital of the PLA Rocket Force.Designed primes and the three SNPs as rs6801836,rs2675511 and rs5182 of AT1R gene were analyzed with allele-specific fluorogenic oligonucleotide probes in an assay combining extension and hybridization.Results The genotype and allele frequencies of the three SNPs were not significantly different between the control group and CAD group (x2 =0.047～2.226,all P＞0.05).The major haplotype constructed with linkaged rs6801836 and rs5182 was TT.The frequency of every haplotypes showed no significantly difference between the two groups (x2 =0.025～1.020,all P＞0.05).Conclusion The three polymorphisms of AT1R gene studied in this work showed no association with CAD susceptibility.
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Objective To investigate the association between single nucleotide polymorphisms (SNP)and expression levels ofα-adducin(ADD1)gene in coronary artery disease (CAD)patients.Methods Extracted DNA and RNA samples of peripher-al blood white cells from 114 CAD patients and 116 healthy individuals in Jan 2011 to Oct 2013 from the General Hospital of the PLA Rocket Force.SNPs of rs3775067 and rs1263359 mutations in the ADD1 gene were analyzed with allele-specific flu-orogenic oligonucleotide probes combining hybridization.The gene expression levels were analyzed with fluorescence labeled and capillary electrophoresis technology.Results The frequencies of the genotypes and alleles of the two SNPs in the ADD1 gene were not significantly different between the two groups (χ2=0.018~1.317,all P>0.05).The ADD1 gene expression levels of CAD group (0.226±0.284)were obviously higher than that of control group (0.153±0.144,P0.05).Conclusion The elevated ADD1 gene expression level would be risk factor for CAD.The polymorphisms of rs3775067 and rs1263359 had no relevance with CAD susceptibility.
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Objective To establish a method for high throughput screening single nucleotide polymorphism(SNP) by tag microarray,and then apply the method to study the gene SNP which is related to the motor function of normal people.Methods The genes related to motor function were firstly defined,and then 48 SNP loci were determined.The rs numbers of these SNP loci were fingered out from PubMed,and the primers were designed with the software in web site "www.autoprimer.com".The primer sequences were then downloaded and sent to the biologic corporation for synthesis.After being synthesized and purified by HPLC these primers were used in the experiments according to the instruction of Bakeman's SNPstream machine.The key techniques of SNPstream machine were tag microarray and single nucleotide extension assay.Once the determination was finished,both the gene frequency and allele frequency of every locus could be statistically analyzed.Results The information of the 48 SNP loci that related to motor function had been determined simultaneously by tag microarray,regardless the number of samples to be detected at the same time.The number of the samples was variable to meet the need.The data of gene frequency and allele frequency of these 48 SNP loci may be used in the subsequent studies.Conclusions Tag microarray used to high throughput screening SNP has the advantages of accuracy,speed,efficiency and reasonable cost.Therefore it can be applied to study the relationship between the SNP and many kinds of diseases.