一种利用基因表达变化检测、鉴别抗生素的新方法

Construction of a promoter reporter array for antibiotic screening

摘要:

本研究利用基因表达变化构建一个高通量检测筛选抗生素化合物的新方法。采用含luxCDABE报道子的质粒 pCS26和pMS402为载体，构建大肠杆菌重组质粒、筛选铜绿假单胞菌以及沙门氏菌的随机启动子库，获得对抗生素有反应的启动子-报道子融合体，构成抗生素筛选体系。通过重复实验筛选出15个启动子-报道子融合体，它们对不同抗生素有不同的反应，通过这一系统可以检测样品中的抗生素，并初步判断与现有抗生素的异同。使用这种方法可以进行高通量筛选潜在的抗菌物质。

Abstract:

We designed and constructed an antibiotic screening system by using antibiotic responsive genes as reporters. Plasmid pCS26 carrying a promoterless luminescence reporter, luxCDABE, was used as the vector and the promoter regions of antibiotic responsive genes/operons from Escherichia coli were cloned upstream of the lux reporter to form the first part of the screening reporter array. Random promoter library of Salmonella enterica and Pseudomonas aeruginosa were screened for antibiotic responsive clones which consist of the second part of the screening array. The selected final reporter array responded to different antibiotics in distinct patterns and enabled in vivo high-throughput screening for antibiotics. Unknown antibiotics could, in general, be classified by analyzing the response patterns. This screening system is both sensitive and efficient and should prove to be a useful tool for screening new antibiotic compounds.