河南农业科学2011，Vol.40Issue(1)：136-140,5.
PRRSV河南株ORF7基因的克隆表达及条件优化
Cloning and Prokaryotic Expression of the ORF7 Gene of PRRSV Henan Strain
摘要
Abstract
Total RNA was extracted from the cell cultures of two Henan strains of porcine reproductive and respiration syndrome virus (PRRSV), HN07 -1 and HN07 -2,and the ORF7 gene of PRRSV was amplified by RT-PCR using specific primers designed according to the sequence of the nucleocapsid protein (N) gene of IAF - exp91. The PCR products were connected into pMD- 19T vector,cloned and sequenced. The results showed that the ORF7 genes of the two strains were same to each other,and the homology of ORF7 gene with HB- 4(hs),VR - 2332,and BJ -4 was 100％, 93.8％ ,and 93.8％ ,respectively. A prokaryotic expression vector pGEX - 6p - 1 containing ORF7 gene was constructed, transformed into E. coli BL21 and induced with IPTG. SDS- PAGE and Western - blot confirmed that the recombinant protein of 40 kD was successfully expressed in E. coli BL 21.关键词
猪繁殖与呼吸综合征/反转录/克隆/表达/Western-blot/美洲型Key words
PRRSV/ RT-PCR/ Cloning/ Expression/ Western- blot/ American genotype分类
农业科技引用本文复制引用
刘明,王爱萍,张改平,肖治军,郭军庆,乔松林,王丽,史西保,刘运超,游雷鸣..PRRSV河南株ORF7基因的克隆表达及条件优化[J].河南农业科学,2011,40(1):136-140,5.基金项目
国家自然科学基金项目(30730068) （30730068）
