LIU Chao-hong, JIAO Chen-song and CHEN Xin-wen. Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells[J]. Virologica Sinica, 2006, 21(3): 217-220.
Citation:
LIU Chao-hong, JIAO Chen-song, CHEN Xin-wen.
Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells .VIROLOGICA SINICA, 2006, 21(3)
: 217-220.
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摘要
以p90-HCV为模板,通过PCR分别得到全长和截短(2881-3078 bp)ns2基因。将截短ns2基因克隆到pET32a原核表达载体,并在大肠杆菌中获得了Trx-His-NS2C融合蛋白的可溶性高表达。通过His 树脂纯化得到融合蛋白,免疫家兔获得高效价高特异性的抗体。同时构建了含有全长ns2基因的重组腺病毒,利用该重组病毒感染HepG2和293细胞成功的表达23.2kDa的NS2蛋白。本文的研究为进一步开展NS2蛋白的结构与功能研究奠定了良好的基础。
关键词:
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NS2
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HCV
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抗体
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表达
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腺病毒
Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells
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1.
1.State Key Laboratory of Virology, Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
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2.
Wuhan General Hospital of Guangzhou Command. Wuhan 430071,China
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Corresponding author:
CHEN Xin-wen,
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Abstract
The full-length and truncated ns2 gene (2881-3078 bp) were gained through PCR using p90-HCV as template. The truncated ns2 gene was cloned into the prokaryotic expression vector PET32a. The fusion protein Trx-His-NS2C expressed in a soluble form in E.coli, and then was purified by His resin. Highly specific and efficient antibody was produced by immunizing the rabbit with purified fusion protein. In addition to this, recombinant adenovirus containing the full-length ns2 gene was constructed. The NS2 protein was expressed successfully in a size of 23.2kDa by infecting 293 and HepG2 cells with recombinant adenovirus. These results had set a base for the further stulies on the structure and function of the NS2 protein.
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References
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Proportional views
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