广西巴马小型猪内源性反转录病毒整合后传代次数对HEK293细胞中HERV-W表达和变异的影响  

Effects of HEK293-PERV-BM passages on the expression and variation of HERV-W in HEK293 cells

作　　者：马玲 钟华[2] 秦树英 白安斌 陈凤莲 覃绍敏 刘金凤 吴健敏 MA Ling;ZHONG Hua;QIN Shu-ying;BAI An-bin;CHEN Feng-lian;QIN Shao-min;LIU Jin-feng;WU Jian-min(Guangxi Key Laboratory of Veterinary Biotechnology,Guangxi Veterinary Research Institute,Nanning 530001,China;Guangxi University for Nationalities,Nanning 530006,China)

机构地区：[1]广西壮族自治区兽医研究所广西兽医生物技术重点实验室,广西南宁530001 [2]广西民族大学,广西南宁530006

出　　处：《中国兽医学报》2020年第10期1918-1923,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31260613);广西自然科学基金资助项目(2015GXNSFAA139089);广西高校中青年教师基础能力提升计划资助项目(KY2016LX079)。

摘　　要：检测不同代次广西巴马小型猪内源性反转录病毒(PERV)感染HEK293细胞模型(HEK293-PERV-BM)中,HERV-W各基因mRNA和syncytin-1蛋白表达水平,以及HERV-W各基因变异情况,评价PERV的整合对HERV-W的影响。结果显示,与HEK293细胞相比,PERV整合后,不同代次HEK293-PERV-BM中HERV-W gag、pol、env和syncytin-1的mRNA相对表达量出现不同程度的改变,且改变趋势相似,即P10或P20后开始升高,至P30最高;选取相对表达量差别较大代次,检测其syncytin-1蛋白,各代次间syncytin-1蛋白表达量的改变较mRNA的差别小。此外,PERV整合后,在P1~P55中HERV-W结构基因仅发生个别位置的点突变,未见发生固定位置的点突变或基因片段的缺失、插入及重组。本试验为系统评价PERV-BM在异种器官移植中的安全性提供参考。To evaluate the effects of the integration of PERV in HEK293 cells on the expression profile of HERV-W,the mRNA expression of HERV-W gag,pol,env and syncytin-1 genes and the protein expression of syncytin-1,as well as the variation of HERV-W were detected in different passages of HEK293-PERV-BM.The qPCR results showed that the mRNA expression of HERV-W gag,pol,env and syncytin-1 genes varied with the passages of HEK293-PERV-BM.In P10 or P20,the genes of HERV-W began to change,and in P30 reached the highest level.The Western blot assay results showed that the protein expression of syncytin-1 was similar to its mRNA expression,but had less change than mRNA.After analysis on the sequencing results of HERV-W gag,pol and env in HEK293-PERV-BM P1-P55,there were only some points mutation in the genes,but no certain points mutation or deletion,insertion or recombination were observed.These results provide experimental basis for further evaluation of the security of PERV-BM in xenotranplantation.

关 键 词：PERV HERV-W syncytin-1 异种器官移植 

分 类 号：S852.65[农业科学—基础兽医学]