犬瘟热病毒H基因原核表达质粒的构建  

Construction of Prokaryotic Expression Vector of H Gene of Canine Distemper Virus

作　　者：周莉[1] 刘志杰[1] 曾智勇[1,2] 汤德元[1] 李春燕[1] 王彬[1] 张晓杰[1] 甘振磊[1] 王凤[1] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病研究室,贵州贵阳550025

出　　处：《贵州农业科学》2011年第9期174-176,共3页Guizhou Agricultural Sciences

基　　金：贵州大学博士基金项目"犬瘟热病毒贵州株的分离鉴定及主要抗原蛋白编码基因的研究"(X060054)

摘　　要：为H蛋白进一步表达作为诊断用抗原、建立特异的犬瘟热病毒(CDV)抗体检测方法及CDV的预防奠定基础,以已构建含有犬瘟热病毒H基因的pMD18-H质粒为模板,利用合成的特异性引物进行PCR扩增,得到1824bp的目的DNA片段,将所得H基因克隆至经相同双酶切处理后的pET32a(+)原核表达载体中,获得重组质粒pET32a(+)-H。测序结果显示,目的基因插入位置和阅读框正确,成功构建了H基因原核表达质粒。To offer accordance with H protein expression as diagnosis antigen,detect CDV specific antibody and provide the basis for CDV prevention,based on the constructed pMD18-H plasmid containing canine distemper virus N gene,the open reading frame of 1 824 bp was amplified by polymerase chain reaction（PCR） using one pair of primers.H gene and pET32a（＋）were digested with the same procedure,then H gene was cloned into pET32a（＋）.The positive recombinant plasmid pET32a（＋）-H was identified and the sequencing results showed that H gene was in the correct inserted position and open reading frame,it conformed that H gene was cloned into expression vector successfully.

关 键 词：犬瘟热病毒 H基因 原核表达 构建 

分 类 号：S852.655[农业科学—基础兽医学]