贵州医科大学学报

2019, v.44;No.224(05) 552-556

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小鼠FGF21基因克隆及his-mFGF21融合蛋白的诱导表达
The Cloning of Mice FGF21 Gene and Induced Expression of His-mFGF21 Fusion Protein

唐青蓝;李红梅;张礼林;王玉丰;颜礼完;周君;
TANG Qinglan;LI Hongmei;ZHANG Lilin;WANG Yufeng;YAN Liwan;ZHOU Jun;Department of Clinical Laboratory, The Third People's Hospital of Hainan Province;Department of Biochemistry and Molecular Biology, Guizhou Medical University;

摘要(Abstract):

目的:对小鼠FGF21基因进行克隆并诱导表达FGF21融合蛋白。方法:以带有小鼠FGF21的T载体pMD19-T-mFGF21为模板特异性扩增FGF21基因片段,将扩增产物克隆至pET-28a(+),得到pET-28a(+)-mFGF21的重组子,并转化于大肠杆菌BL21(DE3),使用IPTG诱导融合蛋白的表达,纯化后采用Western Blot进行鉴定。结果:pET-28(+)-mFGF21重组质粒构建成功,表达出可溶性的his-mFGF21融合蛋白,经纯化后该蛋白his-mFGF21可与FGF21抗体特异性结合。结论:成功构建pET-28(+)-mFGF21重组子,并表达出his-mFGF21蛋白。
Objective: To clone the gene of mice FGF21 to the expression vector pET-28a(+), express the pET-28a(+)-mFGF21 in E.coli, and obtain the fusion protein of mice FGF21. Methods: To specifically amplificate the gene fragment of mFGF21,which was preserved as pMD19-T-mFGF21 in the early stage of the laboratory, and then clone it into pET-28a(+) to get the recombinant plasmids of pET-28a(+)-mFGF21.The pET-28a(+)-mFGF21 was transformed into E.coli BL21(DE3). The fusion protein induced with IPTG and purified in affinity chromatography, was identified by Western blotting. Results: The recombinant plasmid of pET-28a(+)-mFGF21 was successfully constructed and expressed in a soluble form, and the his-mFGF21 protein had specific interaction with FGF21 antibody after being purified. Conclusion: The pET-28a(+)-mFGF21 recombinant can be successfully constructed and the fusion protein of his-mFGF21 can be expressed.

关键词(KeyWords): 成纤维细胞生长因子;基因克隆;融合蛋白;诱导表达;蛋白纯化
fibroblast growth factor;gene cloning;fusion protein;induced expression;protein purification

Abstract:

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基金项目(Foundation): 贵阳市科技局和贵州医科大学联合基金[筑科合同(2017)5-23号]

作者(Author): 唐青蓝;李红梅;张礼林;王玉丰;颜礼完;周君;
TANG Qinglan;LI Hongmei;ZHANG Lilin;WANG Yufeng;YAN Liwan;ZHOU Jun;Department of Clinical Laboratory, The Third People's Hospital of Hainan Province;Department of Biochemistry and Molecular Biology, Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2019.05.011

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