蛇毒cystatin基因真核表达质粒的构建与表达  

Construction of Recombinant Plasmid Containing Snake Venom Cystatin Gene and Its Expression

作　　者：郑海音[1] 林旭[1] 林建银[1] 

机构地区：[1]福建医科大学分子医学研究中心,福州350003

出　　处：《福建医科大学学报》2006年第5期424-426,共3页Journal of Fujian Medical University

基　　金：国家自然科学基金资助项目(30371747);福建省科技厅重大项目资助项目(2002Y003)

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。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 pcDNA 3.1/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.

关 键 词：蛇毒液类 西司他汀类 质粒 转染 基因表达 

分 类 号：Q78[生物学—分子生物学]