广西眼镜蛇毒中Natrin的克隆及在E.coli中的表达  

Gene Cloning and Expression of Natrin Protein from Naja naja atra in E.coli

作　　者：宋慧[1] 马兴才 周燕[1] 焦杨[1] 周元聪[3] 舒雨雁[4] 

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西南宁第七人民医院,530022 [3]中国科学院上海生化与细胞研究所,上海200031 [4]广西医科大学蛇毒研究所,广西南宁530021

出　　处：《时珍国医国药》2012年第3期513-515,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30270304);广西壮族自治区科学基金资助项目(No.0542043;No.0991127)

摘　　要：目的采用基因工程技术,在大肠杆菌中表达广西眼镜蛇毒中Natrin成熟蛋白。方法将来源于广西眼镜蛇毒的Natrin蛋白的基因克隆至融合表达载体pMAL-c2x中,以N端融合了麦芽糖结合蛋白的形式在大肠杆菌BL21(DE3)通过异丙基硫代半乳糖苷诱导表达Natrin蛋白。超声破菌后,将上清液经Amylose树脂柱上纯化后,收集并冻干Natrin融合蛋白,利用蛋白印迹分析其是否有Natrin抗原活性。结果经蛋白印迹分析可知在50 kD处的条带具有Natrin抗原活性。结论在大肠杆菌中可以表达出Natrin蛋白。Objective To express natrin subunit gene from Naja naja atra（Chinese cobra） in E.coli. Methods Natrin was cloned into a fusion expression vector pMAL-c2x,which carried a N-terminal maltose binding protein.After transforming into E.coli BL21（DE3）,natrin was induced to express by IPTG（Isopropyl-thio-β-D-galactoside）.The expressed product was purified by Amylose chelation affinity chromatography.The expressed product was tested by Western blot. Results The recombinant protein had high immunological activities. Conclusion The natrin gene can be successfully expressed in E.coli.

关 键 词：广西眼镜蛇 蛇毒 Natrin 蛋白表达 

分 类 号：R284.2[医药卫生—中药学]