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首页> 外文期刊>Journal of Clinical Microbiology >Distinct Differences in Repertoires of Low-Molecular-Mass Secreted Antigens of Mycobacterium aviumComplex and Mycobacterium tuberculosis
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Distinct Differences in Repertoires of Low-Molecular-Mass Secreted Antigens of Mycobacterium aviumComplex and Mycobacterium tuberculosis

机译：鸟分枝杆菌复杂和结核分枝杆菌低分子质量分泌抗原库的差异

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Antigens in a 4-week-old culture filtrate (CF) ofMycobacterium avium subsp. avium were separated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by Western blotting. The culture had minimal lysis of bacilli, giving a CF preparation consisting mainly of secreted proteins. Comparison with a similar CF of Mycobacterium tuberculosis with almost no contamination with intracellular proteins showed the presence of cross-reactive antigens homologous to the four components of the antigen 85 complex, as well as MPT32. These were major constituents of the M. avium subsp.avium CF. In addition, there were several low-molecular-mass bands (<15 kDa) in both species that did not cross-react with polyclonal and polyvalent rabbit antibodies in Western blotting. Furthermore, these bands were not detected in corresponding sonicate preparations, indicating high localization indexes, which is typical of soluble secreted proteins. A 14-kDa protein was selected for purification and more detailed characterization. The N-terminal amino acid sequence was determined, and a matching gene was found within the genomic sequence of M. avium subsp. avium which was highly homologous to Rv0455c of M. tuberculosis. The gene encoded a signal peptide typical of secreted mycobacterial proteins. A rabbit antiserum was raised against the purified protein, and the antigen was demonstrated by Western blotting in CFs of M. avium subsp. avium, Mycobacterium aviumsubsp. paratuberculosis, Mycobacterium intracellulare, and Mycobacterium scrofulaceum but was not detected in M. tuberculosis. This is a new example of a highly homologous gene being differentially expressed by different mycobacterial species.
机译：禽分枝杆菌亚种4周龄培养滤液（CF）中的抗原。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中分离 avium ，并通过蛋白质印迹法鉴定。培养物对细菌的裂解极少，可得到主要由分泌蛋白组成的CF制剂。与几乎没有被细胞内蛋白质污染的结核分枝杆菌的相似CF的比较显示，存在与抗原85复合物的四个成分以及MPT32同源的交叉反应抗原。这些是 M的主要组成部分。 avium subsp。 avium CF。此外，在两个物种中都有几个低分子质量带（<15 kDa），它们在Western印迹法中不会与多克隆和多价兔抗体发生交叉反应。此外，在相应的超声制剂中未检测到这些条带，表明高定位指数，这是可溶性分泌蛋白的典型特征。选择了一个14 kDa的蛋白质用于纯化和更详细的表征。确定了N末端氨基酸序列，并且在 M的基因组序列内发现了匹配基因。 avium 子空间 avium 与 M的Rv0455c高度同源。结核病。该基因编码一种典型的分泌分枝杆菌蛋白的信号肽。产生针对纯化蛋白的兔抗血清，并通过蛋白质印迹法在 M的CF中证明抗原。 avium 子空间 avium ，禽分枝杆菌亚种。 副结核，胞内分枝杆菌和阴囊分枝杆菌，但在 M中未检测到。结核病。这是由不同的分枝杆菌物种差异表达的高度同源基因的新例子。

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