The Establishment Of The Chinese Minority Genomic DNA Sample Library And CYP2D6 Polymorphisms In Uighur Population Distribution Law

[Purpose]Drugs are usually and specific targets and produce with the effects of drugs, so the effects of drugs and the drug concentration of target parts have close relations, and target parts of the drug concentration and with blood drug concentration exists dynamic balance.Understanding the individual or group of drug metabolism kinetic parameters can predict target parts of the drug concentration andmake personalized medicine becomes possible. In all kinds of drugs in the kinetic parameters, drug metabolism rate affect blood drug concentration is the main factors, and drug metabolism rate and metabolic way are by specific drug metabolizing enzymes of the decision.Cytochrome oxidase p450s is an important human liver drug metabolic enzymes system family, can metabolism substance, drug of endogenous and exogenous compounds, the present study most drug metabolic reactions in the first I enzyme, their metabolic drugs accounted for about40%of the hepatic metabolism drugs. CYP2D6is P450enzyme cytochrome oxidase which is one of the important members and the first to be found a gene polymorphism drug metabolizing enzymes. That has found CYP2D6has more than70kinds of mutations, participate in metabolic clinically20%-25%of the prescription drugs and CYP2D6genetic polymorphism of great ethnic differences and its polymorphism in different between the distribution of race is diverse.The research is proving that the same dose of the drug in the different RACES, different nationalities, and even different between individual metabolic is different, the metabolic difference is drug toxicity and adverse reaction of the main factors, and drug metabolizing enzymes genetic polymorphisms is these undesirable medical effect of one of the main internal factors.The research in the establishment of Chinese minority genomic DNA samples, the protection of the existing ethnic genetic resources, utilization of Uygur genomic DNA samples, through modern molecular biological techniques, on the drug metabolizing enzyme gene CYP2D6in Uygur population of polymorphism distribution for detection, so as to understand the national drug metabolism level difference, optimization therapeutic dose, to decrease the adverse drug reactions to lay adequate experimental base.[Methods]Minzu University of China in the newly enrolled students, through the questionnaire survey, screening within three generations without exogamy history of minority health adults, in the principle of informed consent, in accordance with the standard clinical blood for blood, through a centrifugal column method for genomic DNA extraction. According to the human genetic resources standard encoding uniform numbers, and by agarose gel electrophoresis and ultraviolet spectrophotometer, excluding breaks and degradation of DNA, eliminating less purity samples, cryopreservation, established Chinese minority genomic DNA samples.At present the CYP2D6exon1-5experiments have been completed, the experimental analysis of the CYP2D6exon6-9mutation and polymorphism distribution. The Uyghur population total genomic DNA as template amplification of CYP2D66-9exon, using direct sequencing of each sample were sequenced, using bioinformatics software for mutation screening and mutation, polymorphism distribution of small scale scan. On this basis, through the use of a point mutation in the PCR experiment method with CYP2D6CDS as template obtained2444-2445insG,2459G> A,4124G> C three mutant loci, and the use of double enzyme, links and transformation experiments built three mutation sites of eukaryotic expression plasmid.[Results]1. Blood samples were collected in1800cases, including the han nationality501cases, uighurs145cases.The database covers38ethnic groups and1494genomic DNA samples, including Korean, Mongolian, Dai, Kazak, Uygur, Miao, Hui, Tibetan, Tujia, Yi, Dong, Qiang, Zhuang, Yao, De'ang, Naxi, Xibe, Bai, Kirgiz, Manchu, Hani, Buyi, Gelao, SHE, Li, Shui, Achang, Han Chinese, Uzbeks, Yugur.2. Frequency analysis:in this study we screened2allelic variants of which have been reported and5novel allelic variants, among them, the reported2allelic variants are CYP2D6*2and CYP2D6*10D, the distribution is10%and15%respectively. The frequencies of the other5novel allelic variants run from5%to10%.3. The point mutation method, using the technology of PCR, pEGFP-N1as the carrier, built the success of CYP2D6CDS and2444-2445insG,2459G> A,4124G> C three mutation sites in eukaryotic expression system.[Conclusion]1.Preliminary framework for China's ethnic minorities DNA sample database has been set up.2.CYP2D6gene Uighur population distribution in China with the gene frequency reported from the other national distribution characteristics of significant differences.3.Construction of the CYP2D6CDS and2444-2445insG,2459G> A,4124G> C three mutation sites in eukaryotic expression system.