Abstract:The purpose of this study was to explore the effect of SigE on drug sensitivity of Mycobacterium smegmatis to five antibiotics (Isoniazid, Rifampin, Ethambutol, Levofloxacin, and Ciprofloxacin) and its' mechanism. sigE gene was amplified and cloned into plasmid pMV261 to construct recombinant plasmid pMV261-SigE. The pMV261-SigE was transferred into M. smegmatis to construct recombinant M. smegmatis expressing SigE (pMV261-SigE/MS). Expression of sigE gene was detected by Western blot. M. smegmatis carrying only plasmid pMV261 (pMV261/MS) was used as control. We found that pMV261-SigE/MS group was more resistance to five antibiotics than control group by Resazurine Microtiter Assay and Colony-Forming Units (CFU). Further, the recombinant bacteria were induced into non-replication persistence phase treated with corresponding higher concentration antibiotic for 48 hours. Resuscitation-promoting factor E (RpfE) of M. tuberculosis was cloned and expressed to enhance resuscitation and growth of persistence phase bacteria. The bacteria in persistence phase were incubated in 7H9 containing 20 ng/mL of RpfE for 2 weeks. The bacterial CFU and most probable number (MPN) were measured to count resuscitation index (RI). RI was used to assess bacterial persistence ability which help us to understand the mechanism of SigE effecting drug sensitivity. Among five antibiotics, RI of pMV261-SigE/MS groups in Isoniazid and Ethambutol were significantly higher than that of the control group. So, SigE can influence the sensitivity of isoniazid and ethambutol by promoting M. smegmatis into the non-replication persistence phase.
李智颖, 卢楠, 魏家玮, 唐弘, 吴宣艳, 庄稀尧, 徐蕾, 杨春, 何永林. Sigma E基因对耻垢分枝杆菌五种药物敏感性的影响及机制研究[J]. 中国人兽共患病学报, 2019, 35(2): 131-137.
LI Zhi-ying, LU Nan, WEI Jia-wei, TANG Hong, WU Xuan-yan, ZHUANG Xi-yao, XU Lei, YANG Chun, HE Yong-lin. Effect and mechanism of Sigma E on drug sensitivity of Mycobacterium smegmatis to five antibiotics. Chinese Journal of Zoonoses, 2019, 35(2): 131-137.
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