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在抗结核药物高通量筛选中表达大肠杆菌β-半乳糖苷酶的重组金分枝杆菌

Recombinant Mycobacterium aurum expressing Escherichia coli beta-galactosidase in high throughput screening of antituberculosis drugs.

作者信息

Srivastava R, Kumar D, Srivastava B S

机构信息

Division of Microbiology, Central Drug Research Institute, Lucknow, India.

出版信息

Biochem Biophys Res Commun. 1997 Nov 26;240(3):536-9. doi: 10.1006/bbrc.1997.7689.

Abstract

Mycobacterium aurum is considered a surrogate of M. tuberculosis and recently has been proposed as test organism in high throughput screening of antituberculosis drugs (3). In this investigation, we suggest use of a recombinant M. aurum expressing E. coli lacZ gene, in which beta-galactosidase production is the reporter system as recently reported by us (6). The assay is based on production of beta-galactosidase in presence of drugs during growth. Enzyme production was inhibited within 4 h by frontline antimycobacterial drugs like streptomycin, rifampicin, isoniazid, ethambutol, ofloxacin, and sparfloxacin at their MICs. The assay could be performed conveniently in 96-well microtiter plate format.

摘要

金色分枝杆菌被认为是结核分枝杆菌的替代菌,最近已被提议作为抗结核药物高通量筛选中的测试菌株(3)。在本研究中,我们建议使用表达大肠杆菌lacZ基因的重组金色分枝杆菌,其中β-半乳糖苷酶的产生作为我们最近报道的报告系统(6)。该测定基于在生长过程中药物存在下β-半乳糖苷酶的产生。在其最低抑菌浓度下,一线抗分枝杆菌药物如链霉素、利福平、异烟肼、乙胺丁醇、氧氟沙星和司帕沙星在4小时内抑制了酶的产生。该测定可以方便地在96孔微量滴定板形式中进行。

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