机构地区: 广西大学生命科学与技术学院广西亚热带生物资源保护利用重点实验室
出 处: 《工业微生物》 2005年第3期10-13,共4页
摘 要: 以弗氏柠檬酸菌(Citrobacter freundii)基因组DNA为模板,通过PCR得到甘油脱水酶(glycerol dehydratase)基因dhaB、dhaC、dhaE,克隆到表达载体pSE380上,得到重组质粒pSE-dhaBCE。将此重组质粒转化到E.coliJM109中,重组菌株SDS-PAGE结果显示有明显的61kD、22kD、16kD三条特异性蛋白条带出现。重组菌株经诱导表达,酶活力为11.59U/mL。 The genes dhaB, dhaC, dhaE encoding glycerol dehydratase of Citrobacter freundii were amplified by PCR. The recombinant plasmid pSE-dhaBCE was constructed by inserting dhaB, dhaC, dhaE genes imo expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaBCE. Glycerol dehydratase was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The deduced products of the dhaBCE genes with calculated molecular masses of 61kDa, 22kDa, 16kDa, respectively, revealed high activity. By comparison, no activity appeared in hosted E. coli JM109.