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Spatial and Temporal Expression of Lactobacillus plantarum Genes in the Gastrointestinal Tracts of Mice

机译:植物乳杆菌基因在小鼠胃肠道中的时空表达

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摘要

Lactobacillus plantarum is a common inhabitant of mammalian gastrointestinal tracts, and L. plantarum strain WCFS1 is a human isolate with a known genome sequence. L. plantarum WCFS1 survives intestinal passage in an active form, and its transit time and transcriptional activities were monitored in 15 BALB/c mice at 2, 4, 6, 8, and 24 h after being fed a single intragastric dose of this organism. Enumeration of viable cells isolated from fecal material revealed that the majority of the L. plantarum inoculum transited the mouse intestine within 4 h after ingestion. Three mice were sacrificed at each time point, and total RNA was isolated from the mouse intestinal compartments (stomach through colon). Quantification of L. plantarum 16S rRNA by quantitative real-time reverse-transcription-PCR revealed that L. plantarum was present at elevated levels in the stomach and small intestine for at least 4 h following ingestion and for over 8 h in the cecum and colon. We also examined the expression of 9 L. plantarum housekeeping genes and 15 L. plantarum in vivo-inducible (ivi) genes previously identified by recombination-based in vivo expression technology to be induced in the mouse gastrointestinal tract. The relative expression levels of the ivi genes increased up to 350-fold in the mouse intestine compared to levels observed for L. plantarum WCFS1 cells grown in a rich laboratory medium. Moreover, several genes displayed intestinal compartment-specific (small intestine versus colon) activities. These results confirm that L. plantarum displays specific and differential responses at various sites along the mammalian intestine.
机译:植物乳杆菌是哺乳动物胃肠道的常见动物,而植物乳杆菌WCFS1是具有已知基因组序列的人分离株。植物乳杆菌WCFS1以活性形式在肠道中存活,并且在喂食单次胃内剂量的这种生物后的2、4、6、8和24小时,在15只BALB / c小鼠中监测了其传播时间和转录活性。从粪便中分离的活细胞的计数表明,大多数植物乳杆菌接种物在摄入后4小时内就通过了小鼠肠道。在每个时间点处死三只小鼠,并从小鼠肠腔(通过结肠胃)分离总RNA。通过定量实时逆转录PCR对植物乳杆菌16S rRNA进行定量分析,结果表明,植物乳杆菌在摄入后至少4小时内在胃和小肠中的含量较高,而在盲肠和结肠中则存在8小时以​​上。我们还检查了之前通过基于重组的体内表达技术鉴定的在小鼠胃肠道中诱导的9株植物乳杆菌看家基因和15株植物乳杆菌体内诱导(ivi)基因的表达。与在丰富的实验室培养基中生长的植物乳杆菌WCFS1细胞所观察到的水平相比,小鼠肠道中ivi基因的相对表达水平增加了350倍。此外,一些基因显示出肠区室特异性(小肠对结肠)活性。这些结果证实植物乳杆菌在沿哺乳动物肠道的各个部位显示特异性和差异性应答。

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