| 稳定表达传染性法氏囊病病毒VP2蛋白突变体细胞系的建立 |
| |
| 引用本文： | 孙静,魏永伟,章晓栋,于涟.稳定表达传染性法氏囊病病毒VP2蛋白突变体细胞系的建立[J].浙江大学学报(农业与生命科学版),2011(5). |
| |
| 作者姓名： | 孙静 魏永伟 章晓栋 于涟 |
| |
| 作者单位： | 浙江大学动物科学学院浙江省动物预防医学重点实验室; |
| |
| 摘 要： | 根据传染性法氏囊病病毒(IBDV)HZ2株VP2基因序列设计1对引物,将该基因成功克隆到真核表达载体pDsRed2-N1中,构建重组质粒pDsRed2-N1-VP2(简称PVP2);以PVP2作为过渡载体,运用PCR定点突变技术,对第279位点和第284位点的氨基酸残基分别或同时进行突变,共构建P279、P284和PDA 3株突变体;在此基础上构建能稳定表达HZ2株IBDV及其突变体VP2蛋白的Vero细胞系,间接免疫荧光试验检测表明,Vero细胞系中IBDV VP2蛋白已成功表达.这为进一步深入研究IBDV毒力及抗原变异致病机制奠定了基础;此外,利用定点突变技术改造VP2蛋白,建立不同毒力的突变体,在探求高效、廉价的IBDV DNA疫苗上也有广阔的应用前景和重要意义.
|
| 关 键 词： | 传染性法氏囊病病毒 VP2基因 定点突变 突变体 Vero细胞系 |
Construction of cell line stably expressing IBDV VP2 protein mutants |
| |
| SUN Jing,WEI Yong-wei,ZHANG Xiao-dong,YU Lian.Construction of cell line stably expressing IBDV VP2 protein mutants[J].Journal of Zhejiang University(Agriculture & Life Sciences),2011(5). |
| |
| Authors: | SUN Jing WEI Yong-wei ZHANG Xiao-dong YU Lian |
| |
| Institution: | SUN Jing,WEI Yong-wei,ZHANG Xiao-dong,YU Lian(Zhejiang Province Key Lab of Preventive Veterinary Medicine,College of Animal Sciences,Zhejiang University,Hangzhou 310058,China) |
| |
| Abstract: | A couple of primers was designed on the base of VP2 gene sequence of infectious bursal disease virus(IBDV) HZ2 strain,and the VP2 gene was successfully cloned into eukaryotic expression vector pDsRed2-N1.Then,the recombinant plasmid pDsRed2-N1-VP2 was constructed.The 279 site and 284 site on VP2 protein were mutated singly or homeochronously through site-directed mutagenesis on the constructed vector pDsRed2-N1-VP2.By this way,three mutants of P279,P284 and PDA were constructed in next step.The Vero cell li... |
| |
| Keywords: | infectious bursal disease virus VP2 gene site-directed mutagenesis mutant Vero cell line |
| 本文献已被 CNKI 等数据库收录！ |
|
