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
出处
《工业微生物》
CAS
CSCD
北大核心
2005年第2期19-23,共5页
Industrial Microbiology
基金
辽宁省重大课题(99205002)
