牛副流感病毒3型核衣壳蛋白基因的真核表达  

Eukaryotic expression of nucleocapsid protein gene from bovine parainfluenza virus type 3

作　　者：师新川[1] 温永俊[1] 王凤雪[1] 王炜[1] 王建科[1] 程世鹏[1] 武华[1] 

机构地区：[1]中国农业科学院特产研究所,吉林长春130122

出　　处：《畜牧与兽医》2012年第11期1-4,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家"863"项目(2011AA10A213)

摘　　要：根据牛副流感病毒3型(BPIV-3)内蒙09株(NM09)全基因组序列(GenBank登录号:JQ063064)设计特异性引物,利用RT-PCR方法扩增出牛副流感病毒3型分离株的核衣壳蛋白(N)基因,通过NheI和NotI限制性内切酶位点亚克隆至真核表达载体pcDNA3.1/Zeo(+),获得真核重组质粒pcDNA3.1-N。采用Superfect转染试剂将重组质粒转染至BSR细胞中,转染后的BSR细胞经间接免疫荧光试验和RT-PCR方法检测,重组质粒pcDNA3.1-N在BSR细胞中能正确表达N蛋白。本研究结果为BPIV-3新型疫苗的研制奠定基础。The aim of this study is to construct an eukaryotic expression plasmid for nucleocapsid protein （N） gene of bovine parainfluenza virus type 3 （ BPIV-3 ）. According to the complete sequence of BPIV-3 NM09 strain （ GenBank accession ： JQ063064 ）, a pair of primers was designed and synthesized. The N gene of BPIV3 was amplified by reverse transcription polymerase chain reaction （RT-PCR）. The N gene was cloned into pcDNA3. 1 （＋） vector using NheI and NotI, and thus the expression plasmid pcDNA3.1-N was successfully construc- ted. Then pcDNA3. 1-N was transfected into BSR cell using lipoplast mediate method and positive results were demonstrated by indirect im- munofluorescence assay （IFA） and RT-PCR. The results showed N gene of BPIV-3 was successfully expressed in BSR cell. The study pro- vided fundamental information for the further study on the prevention of BPIV3 infection and new vaccine development.

关 键 词：牛副流感病毒3型 真核表达 N基因 

分 类 号：S852.65[农业科学—基础兽医学]