犬新孢子虫吉林株SAG1基因真核表达载体的构建  

Construction of an eukaryotic expression vector of SAG1 gene from Neospora caninum Jilin strain

作　　者：栾杨[1] 曹世诺[1] 贾立军[1] 张守发[1] 

机构地区：[1]延边大学农学院动物医学系,吉林龙井133400

出　　处：《畜牧与兽医》2009年第9期37-39,共3页Animal Husbandry & Veterinary Medicine

摘　　要：根据GenBank中已发表的犬新孢子虫SAG1基因序列(AF132217),设计了一对含有Kozak序列、起始密码子、终止密码子、BamHⅠ和EcoRⅠ酶切位点的引物。以含有SAG1基因的重组质粒pMD-18T-SAG1为模板,经PCR扩增获得SAG1基因,利用BamHⅠ和EcoRⅠ酶切该片段,回收含有BamHⅠ和EcoRⅠ酶切位点粘性末端的SAG1基因片段,将此基因片段克隆至相同酶切回收后的pVAX1真核表达载体中,获得重组质粒pVAX1-SAG1,经PCR鉴定、酶切分析和重组质粒的序列测定、比较,证实了重组质粒的正确性。Based on the SAG1 gene sequence of Neospora caninum in the GenBank （AF132217）, a pair of primers containing Kozak sequence, initiation codon, termination codon, and BamH Ⅰ and EcoR Ⅰ enzyme digestion sites were designed. Using the recombinant plasmid pMD-18T-SAG1 containing SAG1 gene as template, the SAG1 gene was amplified by PCR. The PCR product was digested with BamH Ⅰ and EcoR Ⅰ , and ligated into the digested eukaryotic expression vector pVAX1 with the same restriction endonuclease. Finally the recombinant plasmid named pVAX1-SAG1 was obtained. Further verification was performed by enzyme digestion, PCR amplification and sequencing.

关 键 词：犬新孢子虫 SAG1基因 真核表达载体 

分 类 号：S852.7[农业科学—基础兽医学]