狂犬病病毒CVS株G基因和IFN-α基因的克隆及双基因真核表达载体的构建  

Cloning and Sequence Analysis of RV CVS Strain Gene and IFN-α Gene and Construction of pIRES-G/IFN-α

作　　者：李江涛[1] 殷相平[1] 柳纪省[1] 李宝玉[1] 李学瑞[1] 杨彬[1] 兰喜[1] 胡永浩[2] 

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农业部畜禽病毒学重点开放实验室,甘肃兰州730046 [2]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《动物医学进展》2007年第9期11-14,共4页Progress In Veterinary Medicine

基　　金：国家高技术研究发展计划863项目(2006AA10A204);甘肃省科技攻关项目(2GS0S2-A41-006-02)

摘　　要：从狂犬病病毒CVS毒株致死的小鼠脑组织中提取总RNA,通过RT-PCR获得G基因;以pMD18-T-α质粒为模板,PCR扩增得到IFN-α基因。两个基因和pIRES真核表达载体分别双酶切,连接构建p IRES-G/IFN-α,经PCR、双酶切鉴定和测序证明载体构建成功。测得的G基因序列长1 575 bp,与CVS株G基因氨基酸同源性为98.5%;IFN-α序列长为570 bp,与GenBank中登录号为NM 001017411的IFN-α基因氨基酸同源性为91.6%。G gene of Rabies virus CVS strain was amplified by RT-PCR and was found to be 1 575 bp in length, the identity of the deduced amino acid sequence of G gene between RV CVS strain and AJ 506997 from GenBank is 98.5%. meanwhile, IFN-α was amplified by PCR and the the identity of deduced amino acid sequence of IFN-α between this PCR product and NM 001017411 from Genbank is 91.6%. G gene was digested with Xho Ⅰ and Mlu Ⅰ and inserted into the cloning site of. p IRES,previously digested with the same enzymes,This recombinant plasmid was designated p IRES-G. IFN -α gene was digested with Sal Ⅰ and Not Ⅰ and ligated with recombinant plasmid p IRES-G, previously digested with the same enzymes, This recombinant plasmid was designated plRES-G/IFN-α.

关 键 词：狂犬病痛毒CVS毒株 G基因 Α干扰素 pIRES载体 

分 类 号：Q786[生物学—分子生物学] S852.659.5[农业科学—基础兽医学]