猪繁殖与呼吸综合征病毒2b基因的克隆和原核表达

摘要

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 26 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.%根据GenBank中猪繁殖与呼吸综合征病毒(PRRSV)美洲株26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222 bp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35 ku,与预期结果相同,且具有良好的免疫学活性.