兔出血症病毒ZB分离株VP60基因克隆、截短表达与活性鉴定  

Cloning,Truncated Expression and Activity Identification of Rabbit Hemorrhagic Disease Virus VP60 Gene of ZB Strain

作　　者：杨丽梅[1,2] 马力[1,2] 张志美[1,2] 徐倩倩[1,2] 郭时金[1,2] 沈志强[1,2] 李峰[1,2] 张颖[1,2] 王艳萍[1,2] 

机构地区：[1]山东省滨州畜牧兽医研究院 [2]山东绿都安特动物药业有限公司,山东滨州256600

出　　处：《动物医学进展》2013年第9期57-61,共5页Progress In Veterinary Medicine

摘　　要：为分析兔出血症病毒地方分离株的变异情况，并获得活性良好的兔出血症病毒基因工程抗原，对兔出血症病毒ZB分离株的VP60基因进行了克隆、序列分析与原核表达。ZB株分离病毒与GenBank中的6株RHDVVP60基因的核苷酸序列同源性在92．5％～97．9％之间，参照ZB株分离病毒VP60基因序列，设计合成一对特异性引物，PCR扩增了876bp的VP60基因片段。将目的片段定向克隆至pET30a表达载体中，经鉴定正确后，重组质粒转化BL21表达菌，经IPTG诱导后获得了以包涵体形式表达的重组蛋白，重组蛋白纯化后，Westernblot检测表明具有良好的抗原性与特异性。In order to analysis variation of rabbit hemorrhagic disease virus isolate, and obtain good active of gene engineering antigens, the research conducted the sequence analysis and prokaryotic expression of rab- bit hemorrhagic disease virus ZB strain VP60 gene. The isolated ZB viruses compared with 6 strains of RHDV VP60 gene in GenBank,the homology of the nucleotide sequence is 92.5% to 97.9%. According to ZB strain VP60 gene sequence, one pairs of primers were designed, 876 bp fragment of VP60 gene was am- plified by PCR. The fragment was directionally cloned into pET30a expression vector. Aafter identifica- tion, the recombinant plasmid was transformed into BL21 expression bacteria and induced by IPTG, the recombinant protein was expressed in inclusion bodys forms, the recombinant protein was purified. West- ern blot detection showed that the expressed protein had good antigenicity and specificity.

关 键 词：兔出血症病毒 VP60基因 克隆 截短表达 

分 类 号：S852.651[农业科学—基础兽医学] Q786[农业科学—兽医学]