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.
出处
《动物医学进展》
CSCD
北大核心
2013年第9期57-61,共5页
Progress In Veterinary Medicine
关键词
兔出血症病毒
VP60基因
克隆
截短表达
Rabbit haemorrhagic disease virus
VP60 gene
cloning
truncated expression
