犬瘟热病毒F蛋白基因片段的克隆与表达及其表达产物抗原性的初步鉴定  

Cloning,Expression of Canine Distemper Virus F Protein Gene Fragment and Identification for Antigencity of Expressive Products

作　　者：许莎琼[1] 傅志强[2] 华修国[1] 朱建国[1] 刘金明[2] 吴祖立[1] 

机构地区：[1]上海交通大学农业与生物学院,上海201101 [2]中国农科院上海家畜寄生虫病研究所,上海200232

出　　处：《上海交通大学学报（农业科学版）》2006年第1期70-74,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市科委攻关项目(No.034909008)

摘　　要：本研究采用RT-PCR法扩增出犬瘟热病毒标准疫苗株的融合蛋白基因(F基因)片段约1499 bp,克隆到pMD18-T载体,酶切鉴定后用一对特异性的引物扩增约732 bp的F蛋白基因片段。并把该基因定向克隆到原核表达载体pET-28a(+)中,转化到宿主菌BL21(DE3)后进行了表达并纯化了该表达产物,Western印迹显示表达产物有良好的抗原性。RT-PCR was used to amply the fusion （F） gene fragment of Canine Distemper Virus in this study. The product which was about 1499 bp was cloned to pMD18-T vector. Then with a set special primers the F protein gene fragment about 732 bp was amplified. The gene fragment was cloned into pET-28a（＋）vector , and the recombinant plasmid was transformed into E.coli BL21（DE3）. After the recombinant E.coli was induced, the expression protein was puritled. The result of Western-blot showed the recombinant protein had good antigencity.

关 键 词：犬瘟热病毒 F基因 克隆 表达 Westem—blot 

分 类 号：S858.292[农业科学—临床兽医学]