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  

Construction of cell line stably expressing IBDV VP2 protein mutants

作　　者：孙静[1] 魏永伟[1] 章晓栋[1] 于涟[1] 

机构地区：[1]浙江大学动物科学学院浙江省动物预防医学重点实验室,浙江杭州310058

出　　处：《浙江大学学报（农业与生命科学版）》2011年第5期473-478,共6页Journal of Zhejiang University:Agriculture and Life Sciences

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.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 line stably expressing VP2 protein and this mutant were also constructed successfully on the base of P279,P284 and PDA by indirect immunofluorescence test.The recombination plasmid and cell line constructed lay the foundation on the deep research about the role and mechanism of 279 site and 284 site in virulence change and antigenic variation of IBDV,which also play a significant role in exploring more efficient and cheaper IBDV DNA vaccination.

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 

分 类 号：Q78[生物学—分子生物学] S858.31[农业科学—临床兽医学]