| pET15b-pep-1-p27mt的构建、表达及意义 |
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| 引用本文： | 王立林 严世荣 汪炳华 王家宁 黄永章. pET15b-pep-1-p27mt的构建、表达及意义[J]. 药物生物技术, 2005, 12(5): 285-290 |
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| 作者姓名： | 王立林 严世荣 汪炳华 王家宁 黄永章 |
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| 作者单位： | 武汉大学医学院，郧阳医学院附属人民医院，武汉大学医学院，郧阳医学院附属人民医院，郧阳医学院附属人民医院 湖北武汉430000，郧阳医学院附属人民医院，湖北十堰442000，湖北十堰442000，湖北武汉430000，湖北十堰442000，湖北十堰442000 |
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| 基金项目： | 湖北省教育厅资助(编号:Q200524001) |
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| 摘 要： | 构建细胞穿膜肽(cell-penetrating peptides,CPPs)PEP-1和细胞周期抑制蛋白p27mt的重组体,并在大肠杆菌中表达。将已构建的表达细胞周期抑制蛋白p27mt的基因,克隆入pET15b-pep-1原核表达载体,在大肠杆菌BL21(DE3)LysS内,以IPTG诱导融合蛋白表达。表达产物用SDS-PAGE及Western blot分析鉴定。结果:成功地构建PEP-1-p27mt融合蛋白原核表达载体,并在IPTG诱导下获得特异性的表达。PEP-1-p27mt融合蛋白表达载体的构建,为体内应用细胞周期抑制蛋白诱导肿瘤细胞凋亡提供了理论基础。
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| 关 键 词： | 细胞穿膜肽 PEP-1 PEP-1-p27mt融合蛋白 pET15b-pep-1-p27mt 重组表达子 抑癌基因 |
| 文章编号： | 1005-8915（2005）05-0285-06 |
| 收稿时间： | 2005-04-06 |
| 修稿时间： | 2005-04-062005-05-25 |
Construction and Expression of pET15b-pep-1-p27mt Recombinant |
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| WANG Li-lin,YAN Shi-rong,WANG Bing-hua,WANG Jia-ning, HUANG Yong-zhang. Construction and Expression of pET15b-pep-1-p27mt Recombinant[J]. Pharmaceutical Biotechnology, 2005, 12(5): 285-290 |
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| Authors: | WANG Li-lin YAN Shi-rong WANG Bing-hua WANG Jia-ning HUANG Yong-zhang |
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| Affiliation: | 1. The Medical College of Wuhan Univercity, Wuhan Hubei 430000, China; 2. Yunyang Medical College, Shiyan Hubei 442000, China |
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| Abstract: | The aim is to construct prokaryotic expression of pET15b-pep-1-p27mt recombinant and to(express) it in E.coli.The cDNAs encoding cyclin dependent kinase inhibitors p27mt was cloned into prokaryotic expression vector pET15b.Recombinant plasmid was transformed into E.coli BL21(DE3)LysS,then was induced with IPTG.The expression of the fusion protein was analyzed by SDS-PAGE and Western blot.The recombinant plasmids were constructed and expressed after IPTG inducting successfully.The obtained PEP-1-p27mt fusion protein has laid the foundation for using cyclin dependent(kinase) inhibitors p27mt in the inducing apoptosis of tumor cells. |
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| Keywords: | Cell-penetrating peptides PEP-1 PEP-1-p27mt fusion protein pET15b-pep-1-p27mt(recombination) expressor Anti-oncogene |
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