【摘要】 目的 :构建可高效生产甘油脱水酶的大肠杆菌工程菌。方法 :将编码甘油脱水酶的三个基因gldA、gldB、gldC ,分别克隆至克隆载体pMD18-T和pSIM -T中 ,经测序正确后 ,再亚克隆至表达融合蛋白的高效表达载体pMAL -c2X上 ,构建成表达质粒pMAL -gldABC ,并转化大肠杆菌E .coliDH5α。结果 :成功地将甘油脱水酶基因gldABC以同向串联方式克隆到大肠杆菌融合表达载体pMAL -c2X中。结论 :得到了含gldABC基因的MBP融合蛋白表达载体 ,为研究甘油脱水酶基因 (gldABC)的在原核表达载体中的串联表达奠定了基础。更多还原

【Abstract】 Objective:To construct the engineered E.coli strain which highly express glycerol dehydratase.Method:The gldA,gldB,gldC gene which encoding glycerol dehydratase α β γ subunit was amplified and cloned into vector pMD18-T and pSIM-T differently.The recombinant plasmid were sequenced with dideoxynucleotide termination method.After the DNA sequence was determined,they were subcloned into E.coli expression vector pMAL-c2X for expressing MBP-gldABC fusion protein.The recombinant plasmid pMAL-gldABC was constructed and transferred into E.coli DH5α.Result:The gldABC gene was successfully cloned into expression vector pMAL-c2X tandem in the same direction.Conclusion:Expression vector pMAL-gldABC for expression MBP-gldABC fusion protein was obtained.This is the basis for expressing gldABC gene in E.coli.更多还原