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Mycobacterium Lrp/AsnC family transcriptional factor modulates the arginase pathway as both a sensor and a transcriptional repressor
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作者 Shuangquan Yan Junfeng Zhen +9 位作者 Yuzhu Li Yu Huang Xuefeng Ai Yue Li Andrea Stojkoska Xue Huang Cao Ruan Jiang Li Lin Fan Jianping Xie 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2021年第11期1020-1031,共12页
L-Arginine is the precursor of nitric oxide(NO),a host immune effector against intracellular pathogens including Mycobacterium tuberculosis(M.tb).Pathogens including M.tb have evolved various strategies targeting argi... L-Arginine is the precursor of nitric oxide(NO),a host immune effector against intracellular pathogens including Mycobacterium tuberculosis(M.tb).Pathogens including M.tb have evolved various strategies targeting arginine to block the production of NO for better survival and proliferation.However,L-arginine metabolism and regulation in Mycobacterium are poorly understood.Here,we report the identification of M.smegmatis MSMEG_1415(homolog of M.tb Rv2324)as an arginine-responsive transcriptional factor regulating the arginase pathway.In the absence of L-arginine,MSMEG_1415 acts as a repressor to inhibit the transcription of the roc(for arginine,ornithine catabolism)gene cluster,thereby switching off the arginase pathway.Treatment with L-arginine relieves the transcriptional inhibition of MSMEG_1415 on the roc gene cluster to activate the arginase pathway.Moreover,the L-arginine-MSMEG_1415 complex activates the transcription of the roc gene cluster by recognizing and binding a 15-bp palindrome motif,thereby preventing the excess accumulation of L-arginine in M.smegmatis.Physiologically,MSMEG_1415 confers mycobacteria resistance to starvation and fluoroquinolones exposure,suggestive of its important role in M.smegmatis persistence.The results uncover a unique regulatory mechanism of arginine metabolism in mycobacteria and identify M.tb Rv2324 as an attractive candidate target for the design of drugs against tuberculosis. 展开更多
关键词 Mycobacterium tuberculosis lrp/asnc family transcriptional regulator REPRESSOR Arginine Arginase pathway Persistence
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结核分枝杆菌Lrp基因(Rv2779c)敲除菌株的构建与功能初探
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作者 郑继芳 尚园园 +2 位作者 鲍生娟 黄海荣 陈素婷 《实用医学杂志》 CAS 北大核心 2023年第19期2428-2433,共6页
目的为深入研究结核分枝杆菌Rv2779c基因的功能,构建针对该基因的敲除突变菌株。方法以结核分枝杆菌H37Rv标准菌株基因组DNA为模板,PCR扩增Rv2779c基因上下游大约500 bp左右的片段作为同源重组的左右臂,并将同源重组左右臂构建到p0004s... 目的为深入研究结核分枝杆菌Rv2779c基因的功能,构建针对该基因的敲除突变菌株。方法以结核分枝杆菌H37Rv标准菌株基因组DNA为模板,PCR扩增Rv2779c基因上下游大约500 bp左右的片段作为同源重组的左右臂,并将同源重组左右臂构建到p0004s穿梭质粒上,将p0004s同源重组质粒构建到phAE159质粒中形成噬菌粒,包装形成的噬菌体侵染H37Rv后将同源重组元件整合到基因组中,通过重组元件中的潮霉素抗性标记对阳性克隆进行筛选,同时利用左右臂扩增及内部片段扩增的原理,对抗性筛选阳性克隆进行鉴定。比较Rv2779c基因敲除突变株与H37Rv标准菌株在正常7H9完全培养基及含高浓度利福平培养基中的生长差异。结果成功构建了包含Rv2779c上下游序列同源重组臂的p0004s-ΔRv2779c重组质粒,并将该质粒与含有分枝杆菌温度敏感型元件的phAE159质粒连接获得重组噬菌粒,最后将包装成功的噬菌体侵染H37Rv获得抗性筛选阳性的克隆菌株,PCR表明结核分枝杆菌Rv2779c基因敲除成功。Rv2779c基因敲除对结核分枝杆菌在正常培养基中生长几乎没有影响,但显著降低了结核分枝杆菌在高浓度利福平培养基中的存活率。结论首次成功构建了结核分枝杆菌Rv2779c敲除菌株,为后续进一步研究Rv2779c基因的功能奠定了重要基础。初步研究发现该基因影响结核分枝杆菌在含利福平培养基中的持留生长。 展开更多
关键词 结核分枝杆菌 基因敲除 lrp/asnc家族蛋白 Rv2779c基因
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Digenic heterozygous mutations in EYS/LRP5 in a Chinese family with retinitis pigmentosa
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作者 Feng-Juan Gao Sheng-Hai Zhang +2 位作者 Jun-Yi Chen Ge-Zhi Xu Ji-Hong Wu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2017年第2期325-328,共4页
Dear Editor,I am Dr.Ji-Hong Wu,from the Department of Ophthalmology,Eye&ENT Hospital of Fudan University,China.I write to present a case report of retinitis pigmentosa(RP)caused by novel digenic heterozygous mutati... Dear Editor,I am Dr.Ji-Hong Wu,from the Department of Ophthalmology,Eye&ENT Hospital of Fudan University,China.I write to present a case report of retinitis pigmentosa(RP)caused by novel digenic heterozygous mutations in a Chinese family. 展开更多
关键词 lrp GENE Digenic heterozygous mutations in EYS/lrp5 in a Chinese family with retinitis pigmentosa
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