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首页> 外文期刊>Infection and immunity >The Largest Open Reading Frame (pks12) in the Mycobacterium tuberculosis Genome Is Involved in Pathogenesis and Dimycocerosyl Phthiocerol Synthesis
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The Largest Open Reading Frame (pks12) in the Mycobacterium tuberculosis Genome Is Involved in Pathogenesis and Dimycocerosyl Phthiocerol Synthesis

机译：结核分枝杆菌基因组中最大的开放阅读框（pks12）参与发病机理和二分枝龙胆酚硫醇的合成。

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The cell wall lipids in Mycobacterium tuberculosis are probably involved in pathogenesis. The largest open reading frame in the genome of M. tuberculosis H37Rv, pks12, is unique in that it encodes two sets of domains needed to produce fatty acids. A pks12-disrupted mutant was produced, and disruption was confirmed by both PCR analysis and Southern blotting. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that a 430-kDa protein band present in the wild type was missing in the mutant. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MS) and liquid chromatography (LC)-MS analysis of tryptic peptides showed that 54 peptides distributed throughout this protein matched the pks12-encoded sequence. Biochemical analysis using [1-14C]propionate as the radiotracer showed that the pks12 mutant was deficient in the synthesis of dimycocerosyl phthiocerol (DIM). SDS-PAGE, immunoblot analysis of proteins, and analysis of fatty acids showed that the mutant can produce mycocerosic acids. Thus, the pks12 gene is probably involved in the synthesis of phthiocerol, the diol required for DIM synthesis. Growth of the pks12 mutant was attenuated in mouse alveolar macrophage cell line MH-S, and the virulence of the mutant in vivo was highly attenuated in a murine model. Thus, pks12 probably participates in DIM production and its expression is involved in pathogenesis.
机译：结核分枝杆菌中的细胞壁脂质可能与发病机理有关。 M基因组中最大的开放阅读框。结核病 H37Rv， pks12 ，其独特之处在于它编码产生脂肪酸所需的两组结构域。产生了 pks12 -破坏的突变体，并通过PCR分析和Southern印迹证实了破坏。十二烷基硫酸钠-聚丙烯酰胺钠凝胶电泳（SDS-PAGE）分析表明，野生型中存在430 kDa的蛋白带，但该突变体中却没有。胰蛋白酶解肽的基质辅助激光解吸电离-飞行时间质谱（MS）和液相色谱（LC）-MS分析表明，分布在该蛋白中的54个肽与 pks12-编码序列匹配。使用[1- 14 C]丙酸酯作为放射性示踪剂的生化分析表明， pks12 突变体在二甲基椰油基苯硫酚（DIM）的合成中存在缺陷。 SDS-PAGE，蛋白质的免疫印迹分析和脂肪酸分析表明，该突变体可产生霉菌酸。因此， pks12 基因可能参与了苯硫酚的合成，苯硫酚是DIM合成所需的二醇。在小鼠肺泡巨噬细胞系MH-S中， pks12 突变体的生长减弱，而在小鼠模型中，该突变体的体内毒性被高度减弱。因此， pks12 可能参与了DIM的产生，其表达也参与了发病机理。

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