荧光假单胞菌M18 rpoD克隆及其对抗生素合成的影响  

Gene Cloning of rpoD and its Impact on Biosynthesis of Antibiotics in Fluorescent pseudomonas M18

作　　者：朱栋华[1] 徐汪节[1] 耿海峰[1] 张雪洪[1] 许煜泉[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海200240

出　　处：《微生物学报》2003年第3期315-323,共9页Acta Microbiologica Sinica

基　　金：国家"十五"科技攻关项目 ( 2 0 0 1BA30 8A0 2 1 4 );上海市科技兴农攻关项目 ( 2 0 0 1 5 1 1 )~~

摘　　要：荧光假单胞菌M1 8对多种植物病原真菌具有显著的抑制作用。荧光假单胞菌(Pseuclomonesfluo rescens)M1 8能同时合成吩嗪 1 羧酸 (PCA)和藤黄绿菌素 (Plt)两种抗生素。从M1 8的基因组中克隆了rpoD基因 ,其相应的氨基酸序列与荧光假单胞菌CHAO中RpoD蛋白的氨基酸序列完全相同。利用基因重组技术和大肠杆菌 荧光假单胞菌穿梭质粒pME60 32 ,将rpoD置于强启动子Ptac的控制下 ,导入M1 8菌株。发现经重组质粒转化的M1 8,与对照相比 ,培养基中PCA和Plt开始累积的时间分别提前 4h和 8h ,积累量提高 1倍和 6倍。Fluorescent pseudomonas M18, one of plant growth promoting rhizobacteria which can inhibit growth of several phytopathogenes, produces several secondary metabolites including antibiotics phenazine-1-carboxylic acid (PCA) and pyoluteorin (Plt). The rpoD gene encoding the housekee- ping sigma factor σ 70 was cloned and sequenced from M18. Through sequencing and homogeneous comparison, the deduced RpoD amino acid sequence between M18 strain and Pseudomonas fluorescens CHAO shows 100% identity. It indicates that rpoD gene is very conserved in different members of fluorescent pseudomonads. The rpoD gene was placed downstream of the constitutive P tac promoter in the shuttle vector pME6032 between E.coli and Pseudomonas fluorescens and the recombinant plasmid was introduced into M18. It was found that the time of both PCA and Plt began to accumulate was 4 and 8 hours earlier and the yield of these antibiotics was increased one and six times more respectively in M18 in comparison with the control.

关 键 词：荧光假单胞菌 M18 克隆 抗生素 合成 rpoD基因 吩嗪-1-羧酸 藤黄绿菌素 

分 类 号：Q785[生物学—分子生物学]