| 蛇毒cystatin基因真核表达质粒的构建与表达 |
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| 引用本文： | 郑海音,林旭,林建银.蛇毒cystatin基因真核表达质粒的构建与表达[J].福建医科大学学报,2006,40(5):424-426. |
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| 作者姓名： | 郑海音 林旭 林建银 |
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| 作者单位： | 1. 福建中医学院中西医结合系
2. 福建医科大学分子医学研究中心,福州,350003 |
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| 基金项目： | 国家自然科学基金;福建省科技厅重大项目资助项目 |
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| 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peDNA3.1/His-cystatin能够在COS7细胞中表达蛇毒cystatin融合蛋白.
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| 关 键 词： | 蛇毒液类 西司他汀类 质粒 转染 基因表达 |
| 文章编号： | 1672-4194（2006）05-0424-03 |
| 收稿时间： | 2006-03-07 |
| 修稿时间： | 2006-03-072006-08-21 |
Construction of Recombinant Plasmid Containing Snake Venom Cystatin Gene and Its Expression |
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| Zheng Haiyin,Lin Xu,Lin Jianyin.Construction of Recombinant Plasmid Containing Snake Venom Cystatin Gene and Its Expression[J].Journal of Fujian Medical University,2006,40(5):424-426. |
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| Authors: | Zheng Haiyin Lin Xu Lin Jianyin |
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| Institution: | Molecular Medicine Research Center of Fujian Medical University, Fuzhou 350004, China |
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| 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 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. |
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| Keywords: | snake venom cystatins plasmid transfection gene expression |
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