一种利用基因表达变化检测鉴别抗生素的新方法-微生物学报.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2010, January 25; 26(1): 93−99 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn ©2010 CJB, All rights reserved. 生物技术与方法 一种利用基因表达变化检测、鉴别抗生素的新方法 赵冰，孙赵麟，杨亮，梁海华，沈立新，段康民 西北大学生命科学学院 西部资源生物与现代生物技术教育部重点实验室，西安 710069 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。 关键词: 抗生素，筛选，启动子，报道子 Construction of a promoter reporter array for antibiotic screening Bing Zhao, Zhaolin Sun, Liang Yang, Haihua Liang, Lixin Shen, and Kangmin Duan Key Laboratory of Resources Biology and Biotechnology in Western China, Ministry of Education, Faculty of Life Sciences, Northwest University, Xi’an 710069, China 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