广西巴马小型猪PERV⁃env基因克隆及其组织表达谱分析

摘要

目的 克隆广西巴马小型猪PERV?env部分基因,并研究其在不同组织中的表达差异.方法 利用RT?PCR方法从巴马小型猪外周血白细胞中扩增PERV?env基因,并与部分国内外已发表的PERV?env基因序列进行同源性及遗传进化分析.以扩增获得的广西巴马小型猪PERV?env基因为模板设计引物,利用半定量RT?PCR的方法对广西巴马小型猪不同组织中PERV mRNA表达情况进行分析.结果 在所检测的9个组织样品中PERV mRNA相对表达水平存在差异,肾及外周血淋巴细胞中PERV mRNA丰度最高,肺、心、肝、脾、胸腺、卵巢次之,且表达丰度差异不显著(P>0.05),胰腺PERV mRNA表达丰度最低.结论 以巴马小型猪胰岛细胞进行异种移植发生PERV感染的潜在危险性可能要低于其它器官,但仍需进一步研究证实.%Objective To clone partial env genes of porcine endogenous retrovirus ( PERV ) isolated from Guangxi Bama minipigs and study their expression differences in different organ tissues. Method The PERV?env genes from peripheral blood leukocytes of Bama minipigs were amplified by RT?PCR, and their homology and phylogenetic analysis were compared with those of some other PERV?env genes published home and abroad. The PERV?env genes obtained by amplification from Bama minipig were used as templates to design primers for analysis on the expressions of PERV mRNA in different tissues of the Bama minipig with a semi?quantitative RT?PCR analysis. Result Differences in the expression levels of PERV mRNA were detected in nine organ tissues of Bama minipig, in which the abundance of PERV mRNA in kidney and peripheral blood lymphocytes was the highest, followed by that in lung, heart, liver, spleen, thymus and ovary, but the expression differences among the six organs were not significant ( P >0. 05 ) , while the expression abundance of PERV mRNA in pancreas was the lowest. Conclusions The potential risk of PERV infection in using the islet cells of Bama mini pig for xenotransplantation is probably lower than using the other organs. However, further research is still needed.