蛇毒液类;_西司他汀类;_质粒;_转染;_基因表达_

蛇毒cystatin基因真核表达质粒的构建与表达

http://www.100md.com 《福建医科大学学报》 2006年第5期

蛇毒液类;,西司他汀类;,质粒;,转染;,基因表达,,蛇毒液类;,西司他汀类;,质粒;,转染;,基因表达,1材料与方法,2结果,3讨论,参考文献

     摘要: 目的构建蛇毒cystatin真核表达载体pcDNA3.1/Hiscystatin,对其在COS7细胞中的表达进行初步研究。方法采用PCR法扩增蛇毒cystatin基因片段,插入pcDNA3.1/His C载体中,测定DNA序列后,转染COS7细胞,利用Westernblot检测COS7细胞中cystatin基因的表达。结果经酶切、测序鉴定证实插入片断已正确,Westernblot检测表明融合蛋白能够在COS7细胞中表达。结论构建的真核表达载体pcDNA3.1/Hiscystatin能够在COS7细胞中表达蛇毒cystatin融合蛋白。

    关键词: 蛇毒液类; 西司他汀类; 质粒; 转染; 基因表达

    Construction of Recombinant Plasmid Containing Snake

    Venom Cystatin Gene and Its Expression

    Zheng Haiyin, Lin Xu, Lin Jianyin

    Molecular Medicine Research Center of Fujian Medical University, Fuzhou 350004, China

    ABSTRACT: Objective To construct recombinant plasmid of snake venom cystatin gene and its expression in the eukaryotic COS7 cells with the vector of pcDNA3.1/HisC. Methods Snake venom cystatin gene was amplified by PCR and inserted into pcDNA3.1/His C. The expression of COS7 cells was detected in by Westernblot assay. Results Cystatin gene was correctly inserted into the vector pcDNA3.1/His C, which was identified by the endonuclease digestion and sequencing. Cystatin can be detected in the plasmid pcDNA3.1/Hiscystatin transfected COS7 cells by Westernblot assay. Conclusion Eukaryotic expression plasmid pcDNA3.1/Hiscystatin was constructed successfully, identifying by it could be expressed in the COS7 cells. ......

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