【作者】 刘鹏； 赵廷昌；

【Author】 Liu peng Zhao Tingchang(Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

【机构】 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室；

【摘要】 本文构建了燕麦食酸菌西瓜亚种(Acidovorax avenae subsp.citrulli)的高效基因组文库,利用群体感应(QS)系统中信号合成基因在Escherichia.coli DH5α的异源表达,以Agrobacterium tumefaciens NTL4(pZLR4)为信号报告菌,从菌株Pslb-94基因组文库中克隆到两个阳性转化子,p5ACC14和p5ACC40。将克隆到的转化子p5ACC14通过电击导入Pseudomonas,fluorescens 2P24△I,成功恢复了信号产生能力。将p5ACC14进一步亚克隆,得到长5.5Kb左右的片段,通过核苷酸序列比对和保守结构域分析,发现其中有1个开放阅读框与NCBI中登陆的A.avenae subsp.citrulli AAC00-1NC008752群体感应基因的核苷酸序列同源性为100%,将获得的基因命名为luxI。构建了luxI基因的原核表达载体,转化Eschefichia coli DH5α,通过报告平板检测,发现有信号产生,进一步说明luxI基因为群体感应信号合成基因。更多还原

【Abstract】 The construction of A.avenae subsp.citrulli genomic library was described here Agrobacterium.tumefaciens NTL4(pZLR4)was used as AHLs reporter strain.The luxl and luxR which were two components of QS system,were cloned from genomic library of Acidovorax avenae subsp citrulli by heterologous expression in E.coli DH5α.In this experiment,The clones p5ACC14 and p5ACC40 from the genomic library was transformed into P.fluorescens 2P24△I by electric shocks,and it recovered the ability of poroducting signals,p5ACC14 was further cloned to get 5.5Kb fragment.It was found that one ORF was related to A.acidovoraxAAC00-1NC008752 luxI with homologous coefficients higher to 100%.We constructed the luxI prokaryotic gene expression vector,transformed it into E.coli DH5α,and detected the product of signal using reporter plate更多还原