Whole-exome Sequencing Identifying Candidate Genes Of Coronary Heart Disease In A Chinese Zhuang Population Family

Background and Objects : Coronary atheroscleroytic heart disease(CAD),short for coronary heart disease(CHD),refers to the pathological and functional changes of coronary artery lead to heart coronary blood vessel lack of oxygen caused by heart disease.At present,the etiology and pathogenesis of coronary heart disease remains unclear.With the deepening of its research,it is found that the genetic effect of coronary heart disease is about 40%-60%,but the significance genetic factors are still unclear.Therefore,studies of the pathogenesis of coronary heart disease has become the current research hotspot.Whole-exome sequencing(WES)is one of the next generation sequencing methods.With the advantages of high sequencing technique and low cost.WES has been the most effective method in disease genetic study.WES is an efficient,cost-effective and efficient method of research,and it was one of the top ten breakthrough in the journal of "Science" in 2010.Up to now,a large number of disease pathogens have been reported hundreds of reports by the application of exome sequencing successfully locate.Therefore,our study analyze the dates of WES sequencing technology to study the candidate genes of coronary heart disease in a family of Guangxi Zhuang population.The study provide the basisfor further study of the pathogenesis of coronary heart disease.Methods:Find out the eligible family of coronary heart disease from 2013 to 2016 in the First Affiliated Hospital of Guangxi Medical University in the Zhuang population.Collected 11 participates from the families about the clinical data and peripheral venous blood 10 ml.Blood,respectively,examine the biochemical and extracted genomic DNA by the rhizosphere DNA extraction kit.The DNA concentration and the DO260/230 value were measured by the nucleic acid protein detector and quality test were tested by agarose gel electrophoresis.Into the sample.The three samples have been carried out by the WES sequencing.The obtained data were controlled by strict quality control,the bioinformatics analysis and the multiple database filtering to narrow the candidate gene range.Then screening out the harmful predicting of amino acid function by using SIFT and Polyphen-2 software.Finally,by the online Mendelian inheritance(OMIM)and NCBI database,analyzing the biological characteristics of candidate genes and the mechanism of coding proteins to screen out Candidate gene of coronary heart disease.The candidate genes were verified by using PCR and Sanger sequencing in the pedigree.Results:The number of SNPs of two patients and one control was 50,and the number of SNPs was 50,and the number of SNPs was 2,and the number of SNPs was 2,The number of points is: 7.After bioinformatics analysis,seven candidate genes were obtained,including four SNPs: RNF25,CCR5,PPP2R3 A,MRPL42,and one was Indel:TMX3.The results of PCR and Sanger sequencing showed that PPP2R3 A and TMX3 were the susceptibility genes for coronary heart diseaseConclusions: The PPP2R3 A and TMX3 genes are highly likely to be susceptibility genes for the familial of coronary heart disease,but whether theyare pathogenic genes for coronary heart disease and subsequent large samples and associated functional validation.