两种组成型启动子PermE*和PSau3A在变铅青链霉菌中的启动效果比较  

Promotion Efficiency Comparison of Constitutive Promoters PermE* and PSau3A in Streptomyces Lividans

作　　者：张莹[1] 陶欣艺[1] 王风清[1] 魏东芝[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室鲁华生物技术研究所,上海200237

出　　处：《化学与生物工程》2013年第11期47-50,54,共5页Chemistry & Bioengineering

摘　　要：红霉素抗性启动子PermE*和来源于链霉菌核基因组片段的PSau3A启动子均属于链霉菌组成型强启动子,分别将它们插入大肠杆菌-链霉菌穿梭质粒pNW-S1的SD序列和转录起始位点之间,构建成两个稳定的组成型表达质粒:pNW-S3和pNW-S4。以磷脂酰丝氨酸合成酶PSS基因为报道基因,以蛋白表达水平评价这两个启动子在变铅青链霉菌TK24中的表达效果。结果表明,在两株组成型表达重组菌中,PSS均得到了高效表达,具有在链霉菌系统中高效表达外源基因的功能,其中PermE*的启动效率略高于PSau3A。Erythromycin resistance promoter PermE＊ and promoter fragment PSau3A from Streptomyces nuclear genome are constitutive promoters stronger than general promoter. They were inserted between the SD sequence and transcription start site of the E. coli-Streptornyces shuttle plasmid pNW-S1 to construct two sta- ble constitutive expression plasmids, pNW-S3 and pNW-S4, phosphatidylserine synthase（PSS） gene, as a re- porter gene, was cloned into plasmid pNW-S3 and pNW-S4 to be transformed into Streptomyces lividans TK24. The promoter activity was evaluated by SDS electrophoresis and PSS activity assay. It showed that PSS was over-expressed with 20 times activity compared to wild strain in TK24, which proved the pNW-S3 and pNW-S4 expression vectors could highly express foreign to that of PSau3A. genes. Promotion effeciency of PermE＊ was superior

关 键 词：组成型启动子 链霉菌 磷脂酰丝氨酸合成酶 

分 类 号：Q785[生物学—分子生物学]