鲨肝再生过程中下调表达基因NT7.1的克隆表达及分离纯化

• 作 者: 袁瑞瑛 叶波平 边杉 吴梧桐
• 机 构: 中国药科大学生命科学与技术学院,江苏,南京,210009
• 出 处: 药物生物技术
• 发表日期: 2009年2期
• 关键词: Shark liver Regeneration Down-regulated gene Prokaryotic expression Separation a

构建了鲨鱼肝再生过程中下调表达基因NT7.1的原核表达载体pET-NT7.1,转化大肠杆菌BL21后,IPTG诱导获得了其原核融合表达产物;利用Hisobind kits对表达产物进行了分离纯化.结果显示,在终浓度0.01 mmol/L的IPTG的诱导下,NT7.1编码的蛋白nt7.1在大肠杆菌中高效表达,目的蛋白占菌体总蛋白的41%,表达产物的相对分子质量大小约为18 000,与预期的相对分子质量大小相当.对于nt7.1的相关研究尤其活性研究仍在进行中,上述工作为研究该未知基因的功能奠定了基础.“,”A novel gene, NT7. 1 (AY599224 in Genbank), which was down regulated during liver regenera-tion, had been identified in regenerated hepatic tissues of shark in our previous work. Since it may play an impor-tant role on the regeneration of Shark liver, uncovering the structure and function of NT7. 1 is an urgent task at present. The prokaryotic expression plasmid pET-NT7. 1 was constructed and transformed into E. coli BL21. Induced with 0. 01mmol/L IPTG, the recombinant protein nt7. I was harvested and purified by affinity chroma-tography, results come out that the quantities of nt7. 1 were about 41% of the total soluble proteins of BL21,while molecular weight was about 18 000Da analyzed by SDS-PAGE electrophoresis, identified with predicted.