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Dynamics of competitive population abundance of Lactobacillus plantarum ivi gene mutants in faecal samples after passage through the gastrointestinal tract of mice

机译:小鼠胃肠道中粪便样品中植物乳杆菌ivi基因突变体竞争种群数量的动态

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摘要

This study aims to evaluate the impact of mutation of previously identified in vivo-induced (ivi) genes on the persistence and survival of Lactobacillus plantarum WCFS1 in the gastrointestinal (GI) tract of mice. Nine Lact. plantarum ivi gene replacement mutants were constructed, focussing on ivi genes that encode proteins with a predicted role in cell envelope functionality, stress response and regulation. The in vitro growth characteristics of the mutants appeared identical to those observed for the wild-type strain, which agrees with the recombination-based in vivo expression technology suggestion that these genes are not transcribed in the laboratory. Quantitative PCR experiments demonstrated differences in the relative population dynamics of the Lact. plantarum ivi mutants in faecal samples after passage through the GI tract of mice. The in situ competition experiments revealed a 100- to 1000-fold reduction of the relative abundance of three of the ivi gene mutants, harbouring deletions of genes predicted to encode a copper transporter, an orphan IIC cellobiose PTS and a cell wall anchored extracellular protein. These experiments clearly establish that the proteins encoded by these three genes play a key role in Lact. plantarum performance during passage of the GI tract.
机译:这项研究旨在评估以前确定的体内诱导(ivi)基因突变对小鼠胃肠道(GI)的植物乳杆菌WCFS1的持久性和存活的影响。九个乳。构建了植物界ivi基因替代突变体,重点研究了ivi基因,该基因编码在细胞包膜功能,应激反应和调节中具有预测作用的蛋白质。突变体的体外生长特性似乎与野生型菌株观察到的相同,这与基于重组的体内表达技术相符,表明这些基因不在实验室中转录。定量PCR实验证明了乳酸菌相对种群动态的差异。通过小鼠胃肠道后粪便样品中的plantarum ivi突变体。原位竞争实验表明,其中三个ivi基因突变体的相对丰度降低了100到1000倍,其中包含预期编码铜转运蛋白,孤儿IIC纤维二糖PTS和细胞壁锚定的细胞外蛋白的基因缺失。这些实验清楚地确定了由这三个基因编码的蛋白质在乳酸中起关键作用。胃肠道通过过程中植物的表现。

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