| PRRSV Hn-1/06株GP5蛋白基因表达载体的构建及高效表达 |
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| 引用本文： | 卢高峰,夏平安,刘明莉,李伟娟,邱璜,李素平.PRRSV Hn-1/06株GP5蛋白基因表达载体的构建及高效表达[J].江西农业学报,2009,21(1). |
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| 作者姓名： | 卢高峰 夏平安 刘明莉 李伟娟 邱璜 李素平 |
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| 作者单位： | 河南农业大学,牧医工程学院,河南,郑州,450002 |
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| 摘 要： | 通过对PRRSV Hn-1/06株ORF5基因序列分析,设计删除信号肽和跨膜功能区的ORF5序列的3对引物,应用重叠延伸PCR以PTG19-T-ORF5质粒为模板进行扩增目的片段,得到长度为410 bp的片段。然后将此基因亚克隆到原核表达载体PET32a,经筛选获得了阳性重组质粒PET32a-ORF5abc,进而在IPTG的诱导下成功表达,获得了大小约为34 kD的融合蛋白GP5abc-His,Western blotting检测结果证实表达的融合蛋白具有良好的生物学活性。
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| 关 键 词： | PRRSV ORF5基因 RT-PCR 原核表达 |
Construction of Prokaryotic Expression Vector and High-level Expression of GP5 Gene of PRRSV Hn-1/06 Strain |
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| LU Gao-feng,XIA Ping-an,LIU Ming-li,LI Wei-juan,QIU Huang,LI Su-ping.Construction of Prokaryotic Expression Vector and High-level Expression of GP5 Gene of PRRSV Hn-1/06 Strain[J].Acta Agriculturae Jiangxi,2009,21(1). |
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| Authors: | LU Gao-feng XIA Ping-an LIU Ming-li LI Wei-juan QIU Huang LI Su-ping |
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| Institution: | College of Animal Husbandry and Veterinary;Henan Agricultural University;Zhengzhou 450002;China |
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| Abstract: | According to analysis of the ORF5 gene order in PRRSV Hn-1/06 strain,three pairs of primers for the deletion of signal peptide and ORF5 sequence in transmembrane domain function area were designed,and PTG19-T-GP5 was used as a template,410 bp segment was obtained by overlap extension PCR amplification.Then after being sub-cloned into the prokaryotic expression vector PET32a,the ORF5abc gene was successfully obtained with the inducement of 1.0 mmol/L IPTG.Western-blotting was performed to confirm that the ex... |
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| Keywords: | PRRSV RT-PCR |
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