甘油脱水酶β亚基融合蛋白在大肠杆菌中的表达和纯化  

Expression and purification of glycerol dehydratase β-subunit in E.coli as fusion protein

作　　者：陈永胜[1] 刘长江[2] 李长彪[2] 翟景波 

机构地区：[1]内蒙古民族大学农学院,内蒙古通辽028042 [2]沈阳农业大学食品学院,辽宁沈阳110161 [3]宝生物工程大连有限公司,辽宁大连116600

出　　处：《工业微生物》2005年第2期19-23,共5页Industrial Microbiology

基　　金：辽宁省重大课题(99205002)

摘　　要：为了表达及纯化甘油脱水酶β亚基蛋白,采用PCR及DNA重组技术将甘油脱水酶β亚基基因gldB重组到含麦芽糖结合蛋白(MBP)的融合蛋白表达载体pMAL-c2x中,在大肠杆菌中进行表达。结果表明,转化重组质粒pMAL/gldB的大肠杆菌经IPTG诱导,SDS-PAGE分析显示:表达出的MBP-gldB融合蛋白相对分子质量约72000,与预期大小一致,并经Westernblot分析证实。进一步通过直链淀粉树脂亲和层析纯化得到电泳均一的融合蛋白。已获得的重组甘油脱水酶β亚基蛋白有利于研究其生物学性能。In order to express and purify the protein subunit of glycerol dehydratase,PCR and DNA recombination techniques were used for cloning gldB gene into expression vector pMAL-c2x and expressing the fusion protein MBP-gldB in E.coli. E.coli DH5α cells with plasmid pMAL/gldB were induced by IPTG for 4 hours. SDS-PAGE analysis showed that the molecular weights of MBP-gldB was about 72000. Western blot analysis indicated that fusion protein presented the specific MBP antigenicity. MBP-gldB fusion protein were purified by amylose resin affinity chromatography and showed to be homogeneity in SDS-PAGE. The MBP-gldB fusion protein obtained was suitable for research of biological functions and potential utility

关 键 词：甘油脱水酶 β亚基融合蛋白 大肠杆菌 原核表达 纯化 1 3-丙二醇 

分 类 号：TQ223.162[化学工程—有机化工]