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目的了解云南地区恒河猴MHC-DRB部分基因的分布情况。方法采用序列特异性引物-聚合酶链式反应(PCR-SSP)分型方法对云南地区4个恒河猴群进行MHC-DRB部分基因检测,用1.5%琼脂糖凝胶电泳检测PCR产物,将有目的片段的部分样品进行测序。结果宁蒗恒河猴群检测出了12个基因,检出率为3.85%~57.69%;昆明自繁恒河猴群检测出了11个基因,检出率为1.63%~42.28%;景东恒河猴群检测出了10个基因,检出率为3.70%~33.33%;镇沅恒河猴群检测出了9个基因,检出率为5.26%~63.16%。测序结果表明,PCR扩增产物的核苷酸序列与GenBank数据库中的序列同源性为92%~99%。卡方检验结果表明,除了MHC-DRB*W201基因外(P<0.05),其余各基因在云南4个恒河猴群体之间的携带率均不存在统计学差异(P>0.05)。结论云南地区恒河猴12个MHC-DRB基因均存在阳性个体,为今后开展具有特定DRB基因的恒河猴群筛选和选育提供了基础,为生物医学相关研究提供合适的具有特定遗传背景的实验动物提供依据。
Objective To understand the distribution of some genes of MHC-DRB in Rhesus macaques in Yunnan Province. Methods The sequence of MHC-DRB gene was detected by PCR-SSP in 4 rhesus monkeys in Yunnan Province. PCR products were detected by 1.5% agarose gel electrophoresis. Part of the target fragment was sequenced. Results Twelve genes were detected in Ningheng rhesus monkeys, and the detection rate was 3.85% -57.69%. Eleven genes were detected in Kunming mimic monkey population, with a detection rate of 1.63% -42.28%. Jingdong Heng The detection rate of 10 genes was 3.70% -33.33% in the group of rhesus monkeys. Nine genes were detected in Zhenyuan rhesus monkeys, with the detection rates ranging from 5.26% to 63.16%. Sequencing results showed that the nucleotide sequence of the PCR product was 92% -99% homologous with the GenBank database. The results of chi-square test showed that there was no significant difference in the carrying rates of other genes between the 4 rhesus macaques in Yunnan (P> 0.05) except MHC-DRB * W201 (P <0.05). Conclusions There are 12 MHC-DRB gene positive individuals in Rhesus monkey in Yunnan Province, which provide the basis for screening and breeding rhesus monkeys with specific DRB gene in the future, and provide a suitable genetic background for biomedical research Experimental animals provide the basis.