Position: Home > Articles > Construction of Neospora caninum ribosomal phosphoprotein gene eukaryotic expression plasmid and transient expression
Animal Husbandry & Veterinary Medicine
2010,42
(9)
30-33
新孢子虫核糖体磷蛋白基因真核表达载体的构建及瞬时表达
作 者：
贾文影;张吉浩;高建伟;杜秋明;许应天;鲁承
单 位：
图们市质量技术监督局;延边大学农学院动物医学系
关键词：
新孢子虫核糖体磷蛋白(NcP0);Vero细胞;真核表达
摘 要：
采用PCR技术扩增出新孢子虫核糖体磷蛋白(NcP0)全长基因片段,并将其克隆入真核表达载体pVAX1中,构建pVAX1-NcP0表达载体。用脂质体法将阳性克隆瞬时转染Vero细胞,对转染细胞进行RT-PCR检测和IFA检测目的基因的转录与表达情况。结果表明克隆的P0蛋白基因序列全长为1 379 bp;RT-PCR结果显示目的基因已被成功转录;IFA检测证明目的基因被成功表达。本试验成功构建了NcP0的真核表达载体,转染Vero细胞,获得了NcP0的瞬时表达。
译 名：
Construction of Neospora caninum ribosomal phosphoprotein gene eukaryotic expression plasmid and transient expression
作 者：
JIA Wen-ying1,ZHANG Ji-hao2,GAO Jian-wei1,DU Qiu-ming1,XU Ying-tian1,LU Cheng1(1.Veterinary Department,Agricultural College of Yanbian University,Longjing 133400,China;2.Tumen Supervisory Bureau for Quality and Technology,Tumen 133100,China)
关键词：
Neospora caninum ribosomal phosphoprotein(NcP0);Vero cells;eukaryotic expression
摘 要：
Full length gene fragment of Neospora caninum ribosomal phosphoprotein(NcP0) was amplified by PCR,and then cloned into the eukaryotic expression vector pVAX1,thus constructing pVAX1-NcP0.The recombinant vector was transfected into Vero cell by lipidosome method.RT-PCR and immunofluoescence assay(IFA) were performed to determine the transcription of the target gene.The results showed that the cloned full length gene was 1 379 bp;RT-PCR demonstrated that the gene has been successfully transcribed;IFA test indicated that the gene was successfully expressed.In the study,the NcP0 eukaryotic expression vector was successfully constructed,and after the vector was transfected into Vero cells,NcP0 obtained the transient expression.
导出引用
计量
文章访问数: 9
HTML全文浏览量: 0
PDF下载量: 1
所属期刊
