猪繁殖与呼吸综合征病毒2b基因的克隆和原核表达  

Cloning and Expression of 2b Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus

作　　者：何斌[1,2,3,4] 李勇[1,2,3,4] 赵建增 高英杰[1] 夏志平[2] 王泽岩[1] 刘雯[1] 张宁[1] 屈雪琪[1] 白雪[1] 梅林[1] 马德慧[3] 张社民 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]中国人民解放军军事医学科学院军事兽医研究所,吉林长春130062 [3]内蒙古民族大学,内蒙古通辽028042 [4]山西省芮城县家畜家禽疫病防治站,山西芮城044600

出　　处：《中国畜牧兽医》2011年第12期67-70,共4页China Animal Husbandry & Veterinary Medicine

基　　金：运城市科技计划项目

摘　　要：根据GenBank中猪繁殖与呼吸综合征病毒(PRRSV)美洲株2b基因序列,设计合成了1对特异性引物,经RT-PCR从PRRSV中扩增出222bp的片段,并克隆至pMD18-T,连接到pGEX-4T-1表达载体,将阳性重组质粒pGEX-4T-2b转化到BL21(DE3),对诱导后的表达产物进行SDS-PAGE电泳和Western blotting分析,所得重组蛋白的分子质量大小为35ku,与预期结果相同,且具有良好的免疫学活性。A pair of specific primers of porcine reproductive and respiratory syndrome virus（PRRSV） 2b were designed and synthesized according to the published gene sequence of PRRSV from GenBank,and 2b gene of PRRSV was obtained by RT-PCR method.The RT-PCR product was cloned into pMD18-T vector and sequenced.After the positive gene was cloned into the prokaryotic expression vector and designated pGEX-4T-2b,the recombinant plasmid was transformed into host cell BL21（DE3） and induced by IPTG.SDS-PAGE and Western blotting test confirmed that the expressed fusion protein is about 35 ku,which had immunogenically reactive activity.

关 键 词：猪繁殖与呼吸综合征病毒 2b蛋白 克隆 表达 

分 类 号：Q78[生物学—分子生物学]