应用毛发PCR法检测猕猴TRIM5基因序列CypA转座子插入的TRIMCyp嵌合基因型

PCR detection of TRIMCyp genotype in Macaques from hair follicle DNA

为检测猕猴TRIM5基因序列CypA转座子插入TRIMCyp突变基因型个体,本研究建立了应用毛发PCR方法,在基因组水平初步检测猕猴属中TRIMCyp基因型方法,并从猕猴属食蟹猴中初步筛选出TRIMCyp突变基因杂合和纯合子个体。实验以平顶猴为阳性对照,无CypA插入的PCR扩增产物片段大小约为2 343 bp,纯合体为3 110 bp,而杂合突变体则为2 343 bp和3 110 bp。与从血液中提取基因组方法检测结果一致。该方法的建立,为进一步调查TRIMCyp嵌合基因型猕猴在中国的分布,以及研究逆转录病毒跨种间传播和建立HIV-1易感猕猴属动物模型提供了实验手段和平台。

A PCR-based method was developed to detect the TRIMCyp genotypes in macaques from genomic DNA extracted from hair follicle.This method could specifically amplify PCR products of 2 343 bp for TRIMCyp homozygote,2 343 bp and 3 110 bp for heterozygote from genomic DNA of Macaca fascicularis,Macaca nemestrina and Macaca mulatta.It was highly reproducible regardless of genomic DNA extracted from hair follicle or from blood leukocyte cells.The application of hair follicle genomic PCR was a sensitive and easy-to-use approach for investigation of the distribution of TRIMCyp in different species of genus Macaca.Monkeys with unique genotype was important models for studies on cross-species infection of retroviruses and on the development of novel nonhuman primate models for HIV-1.