破伤风毒素重组质粒的构建及应用  

Construction and Application of Recombinant Plasmid with Tetanus Toxin Gene

作　　者：姜昌丽[1] 谭德勇[2] 滕毅[1] 马洁[2] 瞿良[1] 程罕松[3] 王惠萱[1] 

机构地区：[1]成都军区昆明总医院检验科,昆明650032 [2]云南大学生物化学与分子生物学实验室,昆明650091 [3]中山大学达安基因股份有限公司,广州510070

出　　处：《中国生物制品学杂志》2010年第11期1214-1216,共3页Chinese Journal of Biologicals

基　　金：863课题子课题:平战时高危感染菌基因实验诊断研究(2006AA020902)

摘　　要：目的构建含破伤风毒素(Tetanus toxin,TT)特异性基因片段的重组质粒,并对其进行应用。方法采用PCR方法,以破伤风灭活菌株基因组DNA为模板,扩增TT的3个特异性基因片段,连接至pMD19-TSimple载体,构建重组质粒pMD19-TTx,以其为阳性参照对模拟破伤风危险品进行PCR检测。结果 PCR及测序证明重组质粒构建正确,以其为阳性参照可快速检出破伤风模拟危险品。结论成功构建了TT重组质粒pMD19-TTx,为破伤风梭菌的检测提供了阳性参照。Objective To construct and apply a recombinant plasmid containing specific tetanus toxin（TT）gene fragment.Methods Three specific TT gene fragments were amplified by PCR using the genomic DNA of inactivated Clostridium tetani strain as a template and linked to vector pMD19-T Simple.Analog tetanus dangerous goods were identified by PCR using the constructed recombinant plasmid pMD19-TTx as a positive reference.Results Both PCR and restriction analysis proved that recombinant plasmid pMD19-TTx was constructed correctly.Using the recombinant plasmid as a positive reference,analog tetanus dangerous goods were detected rapidly.Conclusion Recombinant plasmid pMD19-TTx was successfully constructed,which provided a positive reference for detection of Clostridium tetani.

关 键 词：梭菌 破伤风 破伤风毒素 重组质粒 

分 类 号：Q784[生物学—分子生物学]