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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2008, Vol. 28 Issue (专刊): 21-26    
论文     
DINH与 sC3d3融合基因真核表达质粒的构建及表达
岳耀敬 焦硕 郭宪 冯瑞林
中国农业科学院兰州畜牧与兽药研究所 中国农业科学院兰州畜牧与兽药研究所 中国农业科学院兰州畜牧与兽药研究所 中国农业科学院兰州畜牧与兽药研究所
Construction and expression of DINH and sC3d3 fusion gene eukaryotic expression vector
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摘要:

目的:构建DINH和sC3d3融合分泌型真核表达载体,并在BHK-21细胞中表达。方法: 采用RT-PCR技术扩增绵羊补体C3d基因片段,将该片段插入到pMD18-T载体,然后亚克隆至真核表达载体pcDNA-DPPIS-DINH-mC3d3构建pcDNA-DPPIS-DINH-sC3d3。pcDNA-DPPIS-DINH-sC3d3脂质体法转染BHK-21细胞, Western blotting 鉴定DINH-sC3d3融合蛋白。结果:酶切及测序结果显示, pcDNA-DPPIS-DINH-sC3d3构建正确;在BHK-21细胞中成功地获得了DINH-sC3d3融合蛋白的高效表达。结论: 本实验构建的抑制素基因疫苗可在真核细胞内正确表达,这为进一步的动物实验打下了基础。

Abstract:

Objective: To construct the eukaryotic expression vector for DINH and sC3d3 fusion gene and express it in vitro.Methods: The fragment of C3dwas amplified by RT-PCR and inserted into pMDT-18 plasmid and then sub cloned into eukaryotic expression vector pcDNA-DPPIS-DINH-mC3d3.The recombinant plasmid pcDNA-DPPIS-DINH-sC3d3 was transfected into BHK-21 cells and the expressed product was detected by Western blot. Results: Western blot demonstrated that DINH and sC3d3 existed in the cultural supernatant.Conclusion: The constructed DNA vaccine can be expressed in vitro, which may pave a way for further studies in animals.

收稿日期: 2008-04-08 出版日期: 1900-01-01
通讯作者: 焦硕   
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引用本文:

岳耀敬,焦硕,郭宪,冯瑞林. DINH与 sC3d3融合基因真核表达质粒的构建及表达[J]. 中国生物工程杂志, 2008, 28(专刊): 21-26.

. Construction and expression of DINH and sC3d3 fusion gene eukaryotic expression vector. China Biotechnology, 2008, 28(专刊): 21-26.

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https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2008/V28/I专刊/21

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