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| 结核分枝杆菌H37有毒株与无毒株的代谢相关基因pabB和lpdA启动子活性分析 | |
| 引用本文： | 孙青,吕亮东,姚玉峰,赵英伟.结核分枝杆菌H37有毒株与无毒株的代谢相关基因pabB和lpdA启动子活性分析[J].微生物学报,2010,50(4):459-464. |
| 作者姓名： | 孙青 吕亮东 姚玉峰 赵英伟 |
| 作者单位： | 1. 苏州大学基础医学与生物科学学院,苏州215123;上海交通大学医学院病原生物学教研室,上海200025 2. 复旦大学生命科学学院,上海,200433 3. 上海交通大学医学院病原生物学教研室,上海,200025 4. 苏州大学基础医学与生物科学学院,苏州,215123 |
| 摘 要： | 【目的】通过分析结核分枝杆菌无毒株H37Ra的全基因组序列,并与H37Rv基因组序列比较,发现pabB和lpdA预测的启动子区发生了突变。我们利用报告基因,确认启动子突变与其基因转录水平的关系,探索结核分枝杆菌H37Ra毒力丧失的内在原因。【方法】利用生物信息学方法预测这两对基因的启动子区,采用PCR技术克隆这两对基因的启动子,与分枝杆菌启动子探针载体pMC210相连,DNA测序证实连接片段正确后,用电穿孔法将重组质粒转化至耻垢分枝杆菌mc2155。利用Quantitative Real-Time RT-PCR检测报告基因lacZ转录水平的差异,进一步验证这两对基因启动子的突变对相应基因转录水平的影响。【结果】Quantitative Real Time PCR检测结果显示H37RapabB启动子活性是H37Rv pabB启动子活性的6倍(p0.05),而H37Rv lpdA启动子的活性是H37RalpdA启动子的2倍(p0.05)。【结论】pabB,lpdA的启动子在H37Ra中的突变对其启动子的活性产生了影响,其中lpdA启动子的突变可能与结核分枝杆菌H37Ra的毒力丧失有关。 |
| 关 键 词： | 关键词：结核分枝杆菌 H37Rv H37Ra pMC210 pabB lpdA |
| 收稿时间： | 2009/12/15 0:00:00 |
| 修稿时间： | 2010/1/20 0:00:00 |
Comparative study of the promoter activity of metabolism-related pabB and lpdA genes between Mycobacterium tuberculosis strains H37Rv and H37Ra |
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| Qing Sun,Liangdong Lu,Yufeng Yao and Yingwei Zhao.Comparative study of the promoter activity of metabolism-related pabB and lpdA genes between Mycobacterium tuberculosis strains H37Rv and H37Ra[J].Acta Microbiologica Sinica,2010,50(4):459-464. | |
| Authors: | Qing Sun Liangdong Lu Yufeng Yao and Yingwei Zhao |
| Institution: | Institute of Preclinical Medicine and Biology Sciences, Soochow University, Suzhou 215123;School of Life Sciences, Fudan University, Shanghai 200433;Laboratory of Bacterial Pathogenesis, Department of Medical Microbiology and Parasitology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025;Institute of Preclinical Medicine and Biology Sciences, Soochow University, Suzhou 215123 |
| Abstract: | Objective Comparative genomic study revealed mutations at the promoter regions of pabB and lpdA in avirulent Mycobacterium tuberculosis strain H37Ra compared to its virulent counterpart strain H37Rv.We used LacZ reporter system to test whether those mutations will affect promoter activity and its potential relationship to virulence attenuation of H37Ra.MethodsPromoter regions of pabB and lpdA were predicted by theNeural Network Promoter Predictionmethod(http:/ /www.fruitfly.org /seq_tools /promoter.html).Pr... |
| Keywords: | Mycobacterium tuberculosis H37Rv H37Ra pMC210 pabB lpdA |
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