| 蜂毒素-SP94杂合基因原核表达载体的构建及蛋白纯化 |
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| 引用本文： | 张红绪,曹静,周庆峰,赵鹏,金白洁,王亚威,刘琦,李成伟. 蜂毒素-SP94杂合基因原核表达载体的构建及蛋白纯化[J]. 河南师范大学学报(自然科学版), 2011, 39(3): 122-125 |
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| 作者姓名： | 张红绪 曹静 周庆峰 赵鹏 金白洁 王亚威 刘琦 李成伟 |
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| 作者单位： | 1. 河南师范大学,生命科学学院,河南新乡453007
2. 商丘师范学院,生命科学系,河南商丘476000 |
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| 基金项目： | 国家自然科学基金，河南省教育厅自然科学研究项目，河南省动物学重点学科资助 |
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| 摘 要： | 用可以特异结合到肝癌细胞膜上的小肽SP94,与蜂毒素连接,构建重组表达载体pET32a-蜂毒素-SP94.转入大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE检测表达结果,产物经Ni-NTA Agrosc亲和层析柱纯化后进行Tricine-SDS-PAGE检测.重组蛋白在大肠杆菌中高效表达,约占细胞总蛋白...
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| 关 键 词： | 肝癌 蜂毒素-SP94 载体构建 表达 纯化 |
Construction of Prokaryotic Expression Vector of Melittin-SP94 and Purification |
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| ZHANG Hong-xu,Cao Jing,ZHOU Qing-feng,ZHAO Peng,JIN Bai-jie,WANG Ya-wei,LIU Qi,LI Cheng-wei. Construction of Prokaryotic Expression Vector of Melittin-SP94 and Purification[J]. Journal of Henan Normal University(Natural Science), 2011, 39(3): 122-125 |
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| Authors: | ZHANG Hong-xu Cao Jing ZHOU Qing-feng ZHAO Peng JIN Bai-jie WANG Ya-wei LIU Qi LI Cheng-wei |
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| Affiliation: | 1.College of Life Sciences,Henan Normal University,Xinxiang 453007,China;2.Department of Life Sciences,Shangqiu Normal University,Shangqiu 476000,China) |
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| Abstract: | A small gene SP94 that could specific ally bind to the membrane of hepatoma cell is connected to Melittin,obtaining a fusion gene melittin-SP94.pET32a is chosen as the recombinant vector and pET32a-Melittin-SP94 is constructed,the recombinant protein Trx-Melittin-SP94 is expressed into E.coli BL21(DE3).The recombinant protein is identified by Tricine-SDS-PAGE and purifed by Ni2+ affinity chromatography.The resulting expression of fusion protein Trx-Melittin-SP94 could reach up to 30% of the total cell proteins.More than 50% of the target protein is in a soluble form.High quatified purity protein is obtained by purification.The study has provided a new path of targeted therapy of liver cancer research and clinical applications. |
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| Keywords: | liver cancer melittin-SP94 vector construction expression purification |
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