应用GST pull-down鉴定空肠弯曲菌鞭毛蛋白FlhF和FlhG的相互作用  

Identification of interaction between flagellar related proteins FlhF and FlhG of Campylobacter jejuni by GST pull-down assay

作　　者：王楠 任方哲 宋召军 杨文彬 焦新安 黄金林 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2015年第2期1-4,13,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(31372449);国家高技术研究发展计划项目(863-2012AA101601);国家科技支撑计划项目(2014BAD13B02)

摘　　要：通过构建原核表达载体pCold I-flhG,并在大肠埃希菌(E.coli)BL21中诱导表达带His标签的FlhG和带GST标签的FlhF融合蛋白,应用GST pull-down技术鉴定FlhF和FlhG在体外的结合作用。结果表明:成功构建重组质粒pCold I-flhG,诱导表达获得可溶性的rHis-FlhG和rGST-FlhF蛋白,GST pull-down试验证实FlhF和FlhG之间存在特异性的相互作用,为空肠弯曲菌鞭毛的生物合成以及致病机制研究奠定基础。The prokaryotic expression vector of His fusion flag pCold I-flhG was constructed,then transformed into E.coli BL21 and induced by Isopropylβ-D-1-thiogalactopyranoside（IPTG）to express rHis-FlhG and rGST-FlhF protein.The soluble proteins were analyzed by GST pull-down assay to identify the interaction between the two proteins in vitro.The recombinant expression vector of FlhG was successfully constructed,and the soluble fusion protein rHis-FlhG and rGST-FlhF was obtained by induction and expression.This research confirmed a direct interaction between FlhF and FlhG.The interaction between FlhF and FlhG established a foundation for the flagellar synthesis and pathogenesis of Campylobacter jejuni.

关 键 词：GST pull-down试验 空肠弯曲菌 鞭毛相关蛋白 蛋白间相互作用 

分 类 号：R378.2[医药卫生—病原生物学] Q78[医药卫生—基础医学]