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Comparison of the proteome of Mycobacterium tuberculosis strain H37Rv with clinical isolate CDC 1551

机译:结核菌菌株H37RV蛋白质组的比较临床分离CDC 1551

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The genome sequences of two virulent strains of Mycobacterium tuberculosis (H37Rv and CDC 1551) are now available. CDC 1551 is a recent clinical isolate and H37Rv is a commonly used lab strain which has been subject to in vitro passage. The two strains have been shown to display differing phenotypes both in vivo and in vitro. The proteome of the two strains grown in liquid culture were examined over time to determine whether there are any major differences between them at the protein level and the differences were compared to the genome data. Total cell lysates of the two strains were analysed by two-dimensional electrophoresis. Approximately 1750 protein spots were visualized by silver staining and the protein profiles of the two strains were found to be highly similar. Out of a total of 17 protein spot differences, seven were unique to CDC 1551 and three to H37Rv. Two further spots showed increased intensity in H37Rv, one spot showed differing vertical mobility between the strains and four showed differing spot intensities with time. Twelve of the spot differences were identified using mass spectrometry; however, no obvious association with phenotype could be deduced. When genome differences were analysed and related to the proteome differences, a mobility shift identified in the MoxR protein could be explained by a point mutation at the gene level. This proteome analysis reveals that, despite having been maintained under vastly different conditions, namely in vitro passage and in vivo transmission, these two strains have remained highly similar.
机译:现在可获得两种毒素结核病(H37RV和CDC 1551)的两种毒素菌株的基因组序列。 CDC 1551是最近的临床分离物,H37RV是一种常用的实验室菌株,其已经过体外通过。已经显示出两种菌株在体内和体外显示不同的表型。在液体培养中生长的两种菌株的蛋白质组随着时间的推移,以确定它们在蛋白质水平之间是否存在任何主要差异,并且将差异与基因组数据进行比较。通过二维电泳分析两个菌株的总细胞裂解物。通过银染色可视化约1750个蛋白质点,发现两个菌株的蛋白质谱非常相似。在总共17个蛋白质点差异中,七个是CDC 1551和3至H37RV独特的。两种进一步的斑点在H37RV中表现出增加的强度,一个点在菌株和四个之间显示出不同的垂直迁移率,并且四个显示出不同的点强度随着时间的推移。使用质谱法鉴定了12个点差异;然而,可以推导出与表型无明显关联。当分析基因组差异并与蛋白质组差异有关时,在MOXR蛋白中鉴定的迁移率可以通过基因水平的点突变来解释。该蛋白质组分析表明,尽管在大量不同的条件下维持,即在体外通过和体内传播中,这两个菌株保持高度相似。

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