| PRRSV河南株ORF7基因的克隆表达及条件优化 |
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| 引用本文： | 刘明,张改平,肖治军,郭军庆,乔松林,王丽,史西保,刘运超,游雷鸣,王爱萍.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. |
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| 作者姓名： | 刘明 张改平 肖治军 郭军庆 乔松林 王丽 史西保 刘运超 游雷鸣 王爱萍 |
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| 作者单位： | 1. 郑州大学生物工程系,河南,郑州,450001;河南省农业科学院农业部动物免疫学重点实验室/河南省动物免疫学重点实验室,河南郑州,450002
2. 河南省农业科学院农业部动物免疫学重点实验室/河南省动物免疫学重点实验室,河南郑州,450002 |
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| 摘 要： | 从猪繁殖与呼吸综合症病毒(PRRSV)河南分离株(HN07-1、HN07-2)的细胞培养物中提取RNA,根据参考株IAF-exp91 N蛋白基因序列设计出2对引物,成功克隆出ORF7全长基因,并连接至pMD-19T栽体.基因测序结果表明,2株PRRSV河南分离株ORF7基因序列完全相同;该序列与HB-4(hs)同源性为...
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| 关 键 词： | 猪繁殖与呼吸综合征 反转录 克隆 表达 Western-blot 美洲型 |
Cloning and Prokaryotic Expression of the ORF7 Gene of PRRSV Henan Strain |
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| LIU Ming,ZHANG Gai-ping,XIAO Zhi-jun,GUO Jun-qing,QIAO Song-lin,WANG Li,SHI Xi-bao,LIU Yun-chao,YOU Lei-ming,WANG Ai-ping.Cloning and Prokaryotic Expression of the ORF7 Gene of PRRSV Henan Strain[J].Journal of Henan Agricultural Sciences,2011,40(1):136-140. |
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| Authors: | LIU Ming ZHANG Gai-ping XIAO Zhi-jun GUO Jun-qing QIAO Song-lin WANG Li SHI Xi-bao LIU Yun-chao YOU Lei-ming WANG Ai-ping |
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| Institution: | 1(1.Department of Bioengineering,Zhengzhou University,Zhengzhou 450001,China;2.Key Laboratory of Animal Immunology of the Ministry of Agriculture/Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China) |
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| 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 40kD was successfully expressed in E.coli BL21. |
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| Keywords: | Western-blot |
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