DAB_(389)(Gly_4Ser)_2-αMSH的纯化及细胞特异性毒性  

Investigation of purification and cytotoxicity of DAB_(389)(Gly_4Ser)_2-αMSH

作　　者：李泽鸿[1] 张国利[2] 岳玉环[2] 吴广谋[2] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2007年第6期910-913,918,共5页Chinese Journal of Veterinary Science

基　　金：国家科技部科技型中小企业创新基金资助项目(03C26212200869)

摘　　要：重组表达载体pET28a/DAB389(Gly4Ser)2-αMSH在BL21(DE3)中以可溶形式表达重组蛋白,依次通过硫酸铵盐析、离子交换层析、亲和层析纯化、脱盐得93.26%重组蛋白纯品;细胞活性测试结果表明,该重组毒素只对SP2/0、HeLa、A549、A375等4种癌细胞有杀伤作用,对DK、BHK正常细胞没有杀伤力;重组蛋白对SP2/0、HeLa、A549、A375细胞的IC50分别为:3.138、2.736、7.243、4.672 mg/L。Recombinant plasmid pET28a/DAB389 （Gly4Ser）2-αMSH being transformed into E. coli BL21 （λDE3）, the recombinant toxin was expressed in a soluble protein. The protein was precipitated by ammonium sulfate, DEAE Sepharose Fast Flow,and then purified by Cu Chelating Sepharose Fast Flow,Superdex G-25 chromatogphy, the purity of DAB389 （Gly4Ser）2-αMSH was about 93.26%. DAB389 （Gly4Ser）2-αMSH was proved to be killer on SP2/0,HeLa,A549, A375 cell, and no killability on DK and BHK. The IC50 of the recombinant protein on SP2/0,HeLa,A549,A375 cell were 3. 138,2. 736,7. 243,4. 672 mg/L,respectively.

关 键 词：DAB389(Gly4Ser)2-αMSH 纯化 细胞毒性 

分 类 号：S852.23[农业科学—基础兽医学]