鸡IFN-γ基因重组蛋白复性条件的优化及生物活性鉴定  

Optimization of Refolding Conditions and Bioactivity Identification of cIFN-γ Recombinant Proteins

作　　者：刘鼎阔[1] 王立红[1] 徐大为[2] 杨爱华 王建春 秦操 张勇[1] 

机构地区：[1]鼎正动物药业(天津)有限公司,天津300402 [2]天津农学院,天津300402 [3]天津市动物疫病预防与控制中心,天津300402 [4]新乡市红旗区职业高级中学,河南新乡453200

出　　处：《动物医学进展》2012年第3期52-57,共6页Progress In Veterinary Medicine

基　　金：天津市科技攻关培育计划(05YFGPNC01900);天津市科技计划支撑重点项目(09ZCKFNC00200);天津市科技小巨人周转资金项目(11KQZZNC151)

摘　　要：为提高鸡IFN-γ基因重组蛋白的复性效率,用18种不同的复性缓冲液对在大肠埃希菌中表达的cIFN-γ进行复性处理,通过SDS-PAGE分析、考马斯亮蓝定量、对鸡胚成纤维细胞以及对鸡胚的保护试验检测复性产物的得率和生物学活性。结果表明,去污剂和疏水剂能保证蛋白的复性得率和复性效率,重组表达产物得率可高达94.7%;氧化还原电势是cIFN-γ成功复性的关键,有生物学活性的复性产物的复性液中均存在氧化还原对GSSG/GSH;经去污剂处理后再复性的产物对细胞保护的有效浓度显著低于直接复性的蛋白。18 different refolding buffers were used for renaturation of c-IFN-γ,which is expressed in E.coli.The refolding yield and biological activity of products were detected by SDS-PAGEe,chicken embryo fibroblasts and chicken embryo protection experiments.The results showed that the detergent and the hydrophobic agent ensured protein refolding yield and refolding efficiency,The yield of recombinant product was up to 94.7%;the pair of redox potential-GSSG/GSH,which is the key to successful refolding,existed in refolding solution.The refolding products had biological activity.The effective concentration to protects cells for product by detergent treatment and then refolding is significantly lower than that to direct refolded proteins.

关 键 词：重组鸡γ干扰素 复性条件 生物学活性 

分 类 号：Q784[生物学—分子生物学]