B细胞连接蛋白在原核细胞中的表达和纯化  

Prokaryotic Expression and Purification of BLNK Protein

作　　者：张志英 肖斌[1] 蒋娟 黄秀娜 张蓉[1] 李晓[1] 刘娟子 连菲 杨永泉 张莹莹[1] 孙朝晖 李林海 ZHANG Zhi-Ying;XIAO Bin;JIANG Juan;HUANG Xiu-Na;ZHANG Rong;LI Xiao;LIU Juan-Zi;LIAN Fei;YANG Yong-Quan;ZHANG Ying-Ying;SUN Zhao-Hui;LI Lin-Hai(Guangzhou General Hospital of PLA,Guangzhou 510010,China)

机构地区：[1]中国人民解放军广州总医院,广东广州510010

出　　处：《生物技术通讯》2018年第2期223-226,共4页Letters in Biotechnology

基　　金：广州市产学研协同创新重大专项(20160404003)

摘　　要：目的:利用原核细胞体系表达并纯化B细胞连接蛋白(BLNK)。方法:构建pMAL-c5x-BLNK重组质粒,转化至大肠杆菌BL21(Plyss),IPTG诱导BLNK融合蛋白的表达,通过MBP柱亲和纯化获得目标蛋白,进行12%SDSPAGE分析。结果:构建了BLNK的原核表达质粒pMAL-c5x-BLNK,该质粒在IPTG浓度为0.5 mmol/L、温度为11℃的条件下,可以在大肠杆菌BL21(Plyss)中表达高纯度的BLNK。结论:在原核细胞表达体系中表达并制备了高纯度的BLNK,为下一步研究BLNK的相互作用蛋白和分子结构打下了基础。Objective:To express and purify B-cell linker(BLNK)protein in prokaryotic cell system.Methods:The recombinant plasmid pMAL-c5x-BLNK was constructed and transformed into Escherichia coli BL21(Plyss).BLNK fusion protein was expressed by IPTG inducing.The target protein was obtained by affinity purification with MBP column and analyzed by 12%SDS-PAGE.Results:The prokaryotic expression plasmid pMAL-c5x-BLNK was successfully constructed.Under IPTG concentration of 0.5 mml/L and temperature of 11℃,the recombinant plasmid could express BLNK protein with high purity in E.coli BL21(Plyss).Conclusion:This study successfully constructed prokaryotic expression plasmid of BLNK protein and prepared high purity BNLK protein,which laid a good foundation for further research on the interaction protein and molecular structure of BLNK protein.

关 键 词：B细胞连接蛋白 原核表达 纯化 

分 类 号：Q78[生物学—分子生物学]