抗革兰阴性菌靶向筛选平台的建立  

Establishment of a targeted-specific screening platform against Gram-negative bacteria

作　　者：殷瑜[1,2] 杨天[1,2] 戈梅[1] 

机构地区：[1]上海来益生物药物研究开发中心有限责任公司,上海200240 [2]上海交通大学,上海200240

出　　处：《中国抗生素杂志》2017年第3期183-188,共6页Chinese Journal of Antibiotics

基　　金：国家重大新药创制专项(No.2012ZX09301002-003)

摘　　要：目的建立靶向筛选抗革兰阴性菌新药的综合模型体系。方法以大肠埃希菌DH5α菌株作为革兰阴性菌的模式菌,利用反义RNA沉默技术特异性降低靶基因的表达水平,构建系列反义RNA低表达菌株,并基于这些菌株构建靶向抗菌筛选平台。结果建立了针对72个菌体生长的必需基因,10余个途径的全细胞筛选模型,形成可全面评价化合物活性位点的模型体系。对已知药物进行了模拟筛选和靶点评价,证实了筛选体系的可靠性。结论利用本文建立的筛选体系,可完成抗革兰阴性菌靶向药物筛选和机制评价两个功能。Objective To construct a comprehensive model system for a new antimicrobial discovery targeting Gram-negative bacteria. Methods Eschericia coli DH5α was used as a model strain of Gram-negative bacteria. An array of antisense RNA strains targeting series of essential genes was constructed based on antisense RNA technology. These antisense RNA strains were further used in whole-cell target-specific antibacterial screening platform establishment. Results An array of antisense RNA strains targeting 72 essential genes belong to more than ten metabolic pathways was obtained. These strains were further selected for whole-cell target-specific antibacterial screening platform establishment. A model system for the comprehensive evaluation of the active site of the compound was formed. Known drugs were simulation screened and the target point was evaluated, which confirmed the reliability of the screening system. Conclusion With the screening system established in this paper, two functions of drug screening against G- bacteria and targeted evaluation can be completed.

关 键 词：靶向抗菌筛选平台 革兰阴性菌 反义RNA技术 必需基因 

分 类 号：R978[医药卫生—药品]