| Caspase 3在大肠杆菌E.coli BL21中表达和活性的研究 |
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| 引用本文： | 孙新臣,刘铁刚. Caspase 3在大肠杆菌E.coli BL21中表达和活性的研究[J]. 医学研究生学报, 2005, 18(12): 1059-1062 |
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| 作者姓名： | 孙新臣 刘铁刚 |
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| 作者单位： | 1. 东南大学医学院附属中大医院肿瘤科,东南大学肿瘤研究所,江苏南京,210009
2. 中国医科院医药生物技术研究所肿瘤室,北京,100038 |
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| 基金项目： | 国家自然科学基金资助项目(批准号:30070229);江苏省卫生厅"135工程"开放课题基金资助项目(批准号:K200405);南京市医学发展重点项目基金资助(批准号:ZKX200427) |
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| 摘 要： | 目的：研究凋亡蛋白酶Caspase 3在大肠杆菌中的表达及活性. 方法：用PCR法扩增出Caspase 3基因cDNA,然后插入pCMV-Myc载体,扩增后克隆至表达载体pGEX-6p,使用异丙基硫代-β-D-半乳糖苷（IPTG）诱导Caspase 3蛋白的表达;应用SDS-PAGE和Western blot法鉴定其性质. 结果：经测定诱导3h后Caspase 3活性最高. 结论：可在大肠杆菌E.coli BL21中诱导表达出有活性的Caspase 3,为研究其在细胞凋亡中的作用奠定了基础.
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| 关 键 词： | Caspase 3 基因表达 pGEX-6p 活性测定 |
| 文章编号： | 1008-8199(2005)12-1059-04 |
| 修稿时间： | 2005-08-20 |
The expression and activity of Caspase 3 in E.coli BL21 |
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| SUN Xin-chen,LIU Tie-gang. The expression and activity of Caspase 3 in E.coli BL21[J]. Bulletin of Medical Postgraduate, 2005, 18(12): 1059-1062 |
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| Authors: | SUN Xin-chen LIU Tie-gang |
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| Affiliation: | SUN Xin-chen~ |
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| Abstract: | Objective: To study the expression and activity of the apoptosis protease-Caspase 3 in(E.coli) BL21(DE3). Methods: The cDNA of Caspase 3 was amplified by PCR and inserted into the plasmid pCMV-Myc,it is then cloned to prokaryotic expression vector pGEX-6p,after which Caspase 3 was induced by IPTG.The protein induced was identified by SDS-PAGE and Western blot. Results: After induced by IPTG for 3 hours,the concentration of Caspase 3 reached the highest level. Conclusion: Active Caspase 3 can be induced within E.coli BL21(DE3),further research can be done about the role of Caspase 3 in apoptosis. |
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| Keywords: | Caspase 3 Gene expression pGEX-6p Activity |
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