微生物与感染
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微生物与感染  2016, Vol. 11 Issue (2): 94-99    
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利用表型芯片系统高通量分析H37Ra与H37Rv差异表型
王艺红,郭庆龙,周凤竹,苟宗超,王洪海,张雪莲
复旦大学生命科学学院遗传工程国家重点实验室,上海 200433
High throughput analysis of phenotypic differences between H37Ra and H37Rv using Biolog Phenotype MicroArray
WANG Yihong,GUO Qinglong,ZHOU Fengzhu,GOU Zongchao,WANG Honghai,ZHANG Xuelian
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China
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摘要: H37Rv是结核分枝杆菌标准有毒株,H37Ra是从H37Rv获得的稳定减毒株,但目前H37Ra毒力减弱原因尚不完全清楚。本研究利用表型芯片系统,高通量分析H37Ra生长表型,并与H37Rv表型比较,筛选两菌株表型差异,分析与H37Ra毒力减弱可能的相关表型及分子机制。结果发现,与H37Rv相比,H37Ra耐酸及耐渗透压能力显著下降,且不能利用丁二酸单甲酯和吐温40作为碳源。结核分枝杆菌耐酸能力直接影响其在吞噬体中的生存和代谢,耐高渗能力影响其必需营养物质的跨膜运输,代谢途径的改变影响其在宿主内的能量摄取,三者改变均可能与H37Ra毒力减弱相关。

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王艺红
郭庆龙
周凤竹
苟宗超
王洪海
张雪莲
关键词 结核分枝杆菌表型毒力    
Abstract:Tuberculosis, caused by Mycobacterium tuberculosis (M. tuberculosis) H37Rv, is a human disease of major importance worldwide. The avirulent M. tuberculosis H37Ra which is used commonly in the laboratory was derived from H37Rv in 1935, but the mechanism of its virulence attenuation remains unclear. In order to explore its possible mechanism of virulence attenuation, screening for different phenotypes is important. To make a rapid and high throughput phenotypic analysis between H37Ra and H37Rv, as well as catch the different phenotypes related to virulence attenuation, the Biolog Phenotype MicroArray analysis was conducted. In this research, the phenotypes of H37Ra were determined and compared with its virulent counterpart H37Rv. Of 765 substrates surveyed, H37Ra phenotypes were highly similar to that of H37Rv, but there were several differences in carbon utilization, osmosis and pH arrays. These arrays suggested that H37Ra could not utilize mono-­methyl succinate and Tween 40 as carbon source in contrast to H37Rv. More interestingly, the differences between H37Ra and H37Rv in the tolerance abilities of hyperosmosis and acid were found, and H37Ra could not tolerate circumstance of pH 5.5 or 3% NaCl, but H37Rv could. These metabolic differences may have a disparate impact on the survival of M. tuberculosis in the hosts and be closely related to the virulence attenuation of H37Ra.

Key wordsMycobacterium tuberculosis    Virulence    Phenotype
基金资助:国家自然科学基金(81261120558),上海市科技支撑项目(13431900203、15431900200)

通讯作者: 张雪莲   
引用本文:   
王艺红,郭庆龙,周凤竹,苟宗超,王洪海,张雪莲. 利用表型芯片系统高通量分析H37Ra与H37Rv差异表型[J]. 微生物与感染, 2016, 11(2): 94-99.
WANG Yihong,GUO Qinglong,ZHOU Fengzhu,GOU Zongchao,WANG Honghai,ZHANG Xuelian. High throughput analysis of phenotypic differences between H37Ra and H37Rv using Biolog Phenotype MicroArray. JOURNAL OF MICROBES AND INFECTIONS, 2016, 11(2): 94-99.
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