猪带绦虫六钩蚴TSOL18真核表达载体pVTSOL18m的构建  

Construction of eukaryotic expression vector containing TSOL18 gene of Taenia Solium Oncosphere

作　　者：胡志敏[1] 扈荣良[2] 郑亚东[1] 骆学农[1] 景志忠[1] 岳城[3] 才学鹏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室/甘肃省动物寄生虫病重点研究室,甘肃兰州730046 [2]解放军军事医学科学院军事兽医研究所,吉林长春130062 [3]新疆农业大学动物医学院,新疆乌鲁木齐830052

出　　处：《中国预防兽医学报》2006年第3期302-305,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重大基础研究发展规划(973)项目(No:19990119060)

摘　　要：根据Kozak原理设计引物,以重组原核表达载体pGEX-TSOL18为模板扩增TSOL18基因,用EcoRI、XhoI酶切后与经相同处理的pVAX1载体连接并转化JM109感受态细胞,经测序证明读码框正确后,再根据PCR点突变的方法设计引物,将目的基因NruⅠ位点中的碱基(第347位)经三次PCR反应突变,最终获得含突变位点的TSOL18m基因,EcoRⅠ、XhoⅠ再次酶切后与pVAX1载体连接,成功构建了含点突变的重组pVAX1表达载体,为TSOL18基因重组犬2型腺病毒活载体疫苗的研制奠定了基础。According to Kozak rule, primers which were specific to the open reading frame （ORF） of TSOL18 were designed and the complete TSOL18 fragment was amplified by Polymerase Chain Reaction（PCR）. The amplified gene digesed by EcoR Ⅰ and Xho Ⅰ was ligated into pVAX1 vector, and transformed into JM109 competent cells. Then, according to the method of site mutation by PCR, two pairs of primers were designed and synthesized. After three PCRs, the 1 128 bp mutated fragment was obtained and subcloned into the downstream of CMV promoter of pVAX1 vector,providing the basis of development of live virus vaccine based on TSOL18 gene and canine adenovirus type 2.

关 键 词：猪带绦虫 TSOL18基因 点突变 真核表达载体 

分 类 号：S852.734[农业科学—基础兽医学] S858.28[农业科学—兽医学]