构建同源调节基因的广谱穿梭质粒  

Construction of regulatory genes in broad host range shuttle plasmid

作　　者：晏立英[1] 周乐聪 谈宇俊 单志慧[1] Leonid S.Chernin 沈明珍 

机构地区：[1]中国农业科学院油料作物研究所农业部油料作物遗传改良重点实验室国家油料作物改良中心,湖北武汉430062 [2]以色列耶路撒冷希伯莱大学农学院,rehovot71600

出　　处：《中国油料作物学报》2002年第3期60-62,共3页Chinese Journal of Oil Crop Sciences

基　　金：中以合作项目(SIARF)

摘　　要：利用PCR方法扩增生防细菌成团肠杆菌(Enterobacteragglomerans,IC1270)基因组调节基因GrrS、GrrA、rpoD、rpoS,插入质粒pGEMT-easy中。切下目的片段与酶切穿梭质粒pJFF224-XN连接,经PCR及酶切鉴定,调节基因片段分别以正向或反向插入到质粒的T4启动子下。将带有调节基因的穿梭质粒导入IC1270的感受态细胞,得到带有同源调节基因不同插入方向的IC1270衍生物。抑菌实验结果表明,正向插入的调节基因未提高对病原真菌的生防效果,而反向插入的调节基因丧失对病原真菌的颉抗作用,推测与抑制调节基因的转录有关。Regulatory genes derived from Enterobacter Agglomerans (IC1270) genomic DNA were amplified by PCR, the fragments were inserted into pGEMT-easy vector. The target fragments were cut down from pGEMT-easy and ligated with shuttle vector pJFF224-XN/E.coRI. Through identification, the homogenous regulatory gene inserted in cis and trans direction under T4 promoter, separetely. IC1270 competent cells were transformed with these constructs. The results of the antagonistic assay were: in cis direction, there were no improving biocontrol effect comparing with parent bacteria; in trans direction, derivatives completely lost the antagonistic ability to pathogen, supposed suppressing the transcription of regulatory genes.

关 键 词：调节基因 广谱穿梭质粒 成团肠杆菌 生物防治 原核生物 

分 类 号：S476[农业科学—农业昆虫与害虫防治]