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[应用抑制性消减杂交技术鉴定结核分枝杆菌H37Rv与减毒株H37Ra的差异基因组基因]

[Identification of differential genomic genes of Mycobacterium tuberculosis H37Rv and attenuated strain H37Ra by suppression subtractive hybridization].

作者信息

Xiong Zhi-Hong, Zhuang Yu-Hui, Li Guo-Li

机构信息

Tuberculosis Research Laboratory, 309th Hospital of PLA, Beijing 100091, China.

出版信息

Yi Chuan Xue Bao. 2005 Sep;32(9):979-85.

Abstract

To study the virulence-related genes in Mycobacterium tuberculosis, we used suppression subtractive hybridization to clone the differential genomic genes between Mycobacterium tuberculosis virulence strain H37Rv and attenuated strain H37Ra. All of 54 different genes were cloned, sequenced and analyzed by Southern-blotting. Two different DNA fragments in H37Ra are new genes so far, and get the new Genbank number AY534505 and AY560011. Eight different DNA fragments in H37Rv were obtained. One is the fragment of a gene coding virulence factor mce; one fragment belongs to the gene coding for purC synthenzyme; one for PE family protein; the other 4 fragments for putative gene; and the last one is a non-coding fragment. PCR analysis indicated that 2 of the different genes were present exclusively in the clinical virulent strain and in H37Rv, but not in the clinical avirulent strain and in H37Ra. The novel differential genes may provide an important clue for studying the mechanism of M. tuberculosis pathogenesis.

摘要

为研究结核分枝杆菌中的毒力相关基因,我们采用抑制性消减杂交技术克隆结核分枝杆菌毒力菌株H37Rv和减毒株H37Ra之间的差异基因组基因。共克隆了54个不同基因,并进行测序及Southern杂交分析。H37Ra中的两个不同DNA片段是迄今发现的新基因,获得了新的Genbank登录号AY534505和AY560011。在H37Rv中获得了8个不同DNA片段。其中一个是编码毒力因子mce的基因片段;一个片段属于编码purC合成酶的基因;一个是PE家族蛋白基因;另外4个片段为推测基因;最后一个是无编码功能的片段。PCR分析表明,其中2个不同基因仅存在于临床毒力菌株和H37Rv中,而不存在于临床无毒力菌株和H37Ra中。这些新的差异基因可能为研究结核分枝杆菌致病机制提供重要线索。

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