Use of Mycobacterium smegmatis Deficient in ADP-Ribosyltransferase as Surrogate for Mycobacterium tuberculosis in Drug Testing and Mutation Analysis
Use of Mycobacterium smegmatis Deficient in ADP-Ribosyltransferase as Surrogate for Mycobacterium tuberculosis in Drug Testing and Mutation Analysis
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DOI：
10.1371/journal.pone.0122076
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发表时间：
2015-04-13
期刊：
影响因子：
3.7
通讯作者：
Varshney, Umesh
中科院分区：
文献类型：
--
作者：
Agrawal, Priyanka;Miryala, Sandeep;Varshney, Umesh
利福平是治疗结核病的一线药物。然而，耐药菌株的出现使得合成和测试新的Rif类似物成为必要。耻垢分枝杆菌常被用作M.结核然而，ADP核糖基转移酶（Arr）在M.当与Rif类似物或其它化合物（Rif/Rif类似物）联合使用时，斯美加利汀使Rif失活，使得其不适用于筛选Rif类似物或其它化合物。利福平也用于研究各种DNA修复酶的作用，通过分析RpoB（RNA聚合酶的亚基）突变导致Rif耐药性。当M.以大豆异黄酮为模型。在这里，我们生成了M。通过删除Δ β（Δ β）而获得的smegalciumstrains。分枝Smegalovirus Delta菌株对Rif的最低抑菌浓度（MIC）与M相似。结核异烟肼、吡嗪酰胺、乙胺丁醇、环丙沙星和链霉素对M. smegmatis Delta arr. Delta UdgB和Delta UdgB联合（udgB编码切除尿嘧啶的DNA修复酶）菌株的生长曲线和突变谱与其对应的野生型UdgB相似。然而，Delta fpg Delta fpg菌株的突变谱与Delta fpg菌株的突变谱略有不同（fpg编码切除8-oxo-G的DNA修复酶）。我们的研究表明M.结论：Smegalovirus Delta菌株是DNA修复研究中药物检测和突变谱测定的理想模型系统。
Rifampicin (Rif) is a first line drug used for tuberculosis treatment. However, the emergence of drug resistant strains has necessitated synthesis and testing of newer analogs of Rif. Mycobacterium smegmatis is often used as a surrogate for M. tuberculosis. However, the presence of an ADP ribosyltransferase (Arr) in M. smegmatis inactivates Rif, rendering it impractical for screening of Rif analogs or other compounds when used in conjunction with them (Rif/Rif analogs). Rifampicin is also used in studying the role of various DNA repair enzymes by analyzing mutations in RpoB (a subunit of RNA polymerase) causing Rif resistance. These analyses use high concentrations of Rif when M. smegmatis is used as model. Here, we have generated M. smegmatis strains by deleting arr (Delta arr). The M. smegmatis Delta arr strains show minimum inhibitory concentration (MIC) for Rif which is similar to that for M. tuberculosis. The MICs for isoniazid, pyrazinamide, ethambutol, ciprofloxacin and streptomycin were essentially unaltered for M. smegmatis Delta arr. The growth profiles and mutation spectrum of Delta arr and, Delta arr combined with Delta udgB (udgB encodes a DNA repair enzyme that excises uracil) strains were similar to their counterparts wild-type for arr. However, the mutation spectrum of Delta fpg Delta arr strain differed somewhat from that of the Delta fpg strain (fpg encodes a DNA repair enzyme that excises 8-oxo-G). Our studies suggest M. smegmatis Delta arr strain as an ideal model system in drug testing and mutation spectrum determination in DNA repair studies.