| 丙型肝炎病毒(HCV)截短型E2的原核表达和蛋白纯化 |
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| 引用本文： | 姚敏,方海亮,尹文,雷迎峰,杨敬,孙梦宁,田江红,吕欣. 丙型肝炎病毒(HCV)截短型E2的原核表达和蛋白纯化[J]. 科学技术与工程, 2009, 9(9) |
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| 作者姓名： | 姚敏 方海亮 尹文 雷迎峰 杨敬 孙梦宁 田江红 吕欣 |
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| 作者单位： | 第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032;第四军医大学基础部微生物学教研室,西安,710032 |
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| 摘 要： | 对丙型肝炎病毒(HCV)截短型包膜糖蛋白E2-661基因进行原核表达和纯化.利用PCR法扩增出661 bp的截短型HCV E2区基因片段,并将测序正确的E2-661基因克隆入原核表达载体pET28a中,转化大肠杆菌BL21(DE3),经IPTG诱导表达后,通过SDS-PAGE和Western-blot对表达产物进行分析和鉴定.结果表明目的蛋白分子量约为35 kDa,主要以包涵体形式大量存在.Western-blot结果显示目的蛋白与抗His标签单抗及HCV阳性血清均具有良好的反应原性.并通过镍离子亲和层析方法获得纯化的重组蛋白.以上结果为HCV E2功能的进一步研究奠定了基础.
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| 关 键 词： | 丙型肝炎病毒 包膜糖蛋白E2 原核表达 |
Prokaryotic Expression and Purification of Truncated Hepatitis C Virus Envelope Glycoprotein E2 |
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| YAO Min,FANG Hai-liang,YIN Wen,LEI Ying-feng,YANG Jing,SUN Meng-ning,TIAN Jiang-hong,L Xin. Prokaryotic Expression and Purification of Truncated Hepatitis C Virus Envelope Glycoprotein E2[J]. Science Technology and Engineering, 2009, 9(9) |
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| Authors: | YAO Min FANG Hai-liang YIN Wen LEI Ying-feng YANG Jing SUN Meng-ning TIAN Jiang-hong L Xin |
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| Affiliation: | Department of Microbiology;Fourth Military Medical University;Xi'an 710032;P.R.China |
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| Abstract: | To construct a recombinant prokaryotic plasmid of truncated HCV E2(E2-661)and observe its expression in E.coli,HCV E2-661 gene fragment was amplified by PCR.After verified by sequencing.It was inserted into the prokaryotic expression vector plasmid pET28a.The recombinant plasmid pET28a-E2-661 was then transformed into E.coil BL21(DE3)to express the fusion protein induced with IPTG.The expression product was detected by SDS-PAGE and western-blot.The fusion protein mainly aggregated into the inclusion body.Th... |
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| Keywords: | hepatitis C virus envelope glycoprotein E2 prokaryotic expression |
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