猪繁殖与呼吸综合征病毒BJ-4株ORF3基因的原核表达

通过 PCR方法从重组质粒 p GEM- ORF3扩增得到缺失 N端疏水序列的基因片段 d ORF3(deleting ORF3)。将d ORF3克隆至原核高效表达载体 p GEX- 4 T- 2 ,在 E.coli BL 2 1细胞中成功表达了猪繁殖与呼吸综合征病毒 (PRRSV)重组蛋白GST- d ORF3,表达产物以包涵体的形式存在 ,表达量为 30 .6 % ,Western- Blot结果表明重组蛋白可被 PRRSV阳性血清所识别。表达的重组蛋白为进一步研究 PRRS病毒次要结构蛋白 GP3的免疫特性和功能奠定了基础

Prokaryotic expression of the ORF3 gene of porcine reproductive and respiratory syndrome virus BJ-4

The dORF3 deleting hydrophobic N-terminal sequence was successfully amplified from recombinant plasmids pGEM-ORF3 by PCR and cloned into prokaryotic expression vector pGEX-4T-2. The recombinant fusion protein GST-dORF3 was highly expressed in E. coli BL21 and could amount to 30.6% of the total bacterial proteins. Western-Blot analysis showed the recombinant protein was able to react with porcine polyclonal antibody against PRRSV. The result provided fundamental data and materials for the further investigation on immunogenicity and function of GP3 of PRRSV.