SARS冠状病毒RNA聚合酶基因片段体外组装和siRNA干扰研究  

Assembling the RdRp gene fragment of SARS coronavirus in vitro by assembly PCR and RNA interference test

作　　者：马成川 谭光宏[2] 阳宇[1] 李灵[1] 

机构地区：[1]四川大学生命科学学院功能基因组实验室,生物资源与环境教育部重点实验室,成都610064 [2]海南医学院,海南省热带病重点实验室,海口571101

出　　处：《四川大学学报（自然科学版）》2014年第6期1307-1311,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(31000579)

摘　　要：为了安全便捷地获取严重急性呼吸系统综合征冠状病毒(Severe Acute Respiratory Syndrome coronavirus,SARS coronavirus)的RdRp基因,作者合成了26条寡聚核苷酸片段,利用组装PCR(assembly PCR)在体外构建了SARS病毒的RdRp基因片段,并建立了基于Vero E6细胞的SARS RdRp基因的稳定表达株.之后设计了4对针对SARS RdRp基因的siRNA,基因干扰表明这4对siRNA均能高效干扰SARS RdRp基因在Vero E6细胞中的表达.至此,本文提供了一种安全便捷地获取病毒基因的方法.To get SARS coronavirus from DNA shuffling was established sembled from 26 o stable expression ce NAs against 645bp- showed that all the rent research therefore RdRp gene conveniently, and a 645bp fragment of Rd an assembly PCR proceduredeveloped Rp gene of SARS coronavirus was as- gosin vitro. Thereafter,the 645bp-RdRp was transfected into Vero E6 lls, Vero-RdRp, were selected out for RNA interference test. Four syn RdRp were used to interfere RdRp RNA transcribed from Vero-RdRp. four siRNAs decreased the expression of the assembled RdRp efficient ore provided an efficient strategy to generate the access-limited virus ge cells and the thesized siR- The results ly. The cur- nes in vitro.

关 键 词：SARS冠状病毒 RDRP DNA改组 

分 类 号：Q78[生物学—分子生物学]