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目的探讨建立检测EsD点突变rs9778的片段长度差异等位基因特异性PCR技术(Fragment length discrepant allele specific PCR,FLDAS-PCR),并调查贵州汉族群体遗传学参数。方法针对EsD基因点突变(NCBI Reference SNP ID:rs9778),设计片段长度差异等位基因特异性PCR引物进行扩增,经丙烯酰胺凝胶电泳分离、银染显带后分型,并调查175例贵州省汉族无关群体等位基因数。结果片段长度差异等位基因特异性PCR技术对EsD-rs9778亚型的3种基因型分型明确。贵州省汉族人群EsD-rs9778 A/G基因频率分别为0.651和0.349,基因型频率分别为1/1 0.434、1/2 0.434和2/2 0.132,Ho 0.434、He 0.457、PIC 0.351、DP 0.599、PEt 0.176 PEd 0.103,基因型分布符合HardyWeinberg平衡。结论片段长度差异等位基因特异性PCR技术对EsD-rs9778亚型的3种基因型分型明确,该位点多态性有实际应用价值。
OBJECTIVE: To establish a FLDAS-PCR method for allelic length polymorphism (PCR-RFLP) of rs9778 in EsD loci and to investigate the population genetic parameters of Guizhou Han population. Methods According to the point mutations of the EsD gene (NCBI Reference SNP ID: rs9778), allele-specific PCR primers were designed to amplify the fragment length. The PCR products were separated by acrylamide gel electrophoresis and stained with silver stain. Guizhou Han unrelated population alleles. Results The fragment length difference allele-specific PCR was used to identify the three genotypes of EsD-rs9778 subtype. The frequencies of the A / G gene of EsD-rs9778 in Guizhou Han nationality were 0.651 and 0.349, respectively, and the genotype frequency was 1/1 0.434, 1/2/2344 and 2/2 0.132, Ho 0.434, He 0.457, PIC 0.351, DP 0.599, PEt 0.176 PEd 0.103, genotype distribution in line with Hardy Weinberg equilibrium. Conclusion The allele-specific PCR of fragment length differs from the three genotypes of EsD-rs9778 subtypes. The polymorphism of EsD-rs9778 is of practical value.