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首页> 外文期刊>Applied Microbiology >RNA–Stable-Isotope Probing Shows Utilization of Carbon from Inulin by Specific Bacterial Populations in the Rat Large Bowel
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RNA–Stable-Isotope Probing Shows Utilization of Carbon from Inulin by Specific Bacterial Populations in the Rat Large Bowel

机译:RNA稳定同位素探测显示大鼠大肠中特定细菌群体从菊粉中碳的利用

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Knowledge of the trophisms that underpin bowel microbiota composition is required in order to understand its complex phylogeny and function. Stable-isotope (~(13)C)-labeled inulin was added to the diet of rats on a single occasion in order to detect utilization of inulin-derived substrates by particular members of the cecal microbiota. Cecal digesta from Fibruline-inulin-fed rats was collected prior to (0 h) and at 6, 12, 18 and 24 h following provision of the [~(13)C]inulin diet. RNA was extracted from these cecal specimens and fractionated in isopycnic buoyant density gradients in order to detect ~(13)C-labeled nucleic acid originating in bacterial cells that had metabolized the labeled dietary constituent. RNA extracted from specimens collected after provision of the labeled diet was more dense than 0-h RNA. Sequencing of 16S rRNA genes amplified from cDNA obtained from these fractions showed that Bacteroides uniformis , Blautia glucerasea , Clostridium indolis , and Bifidobacterium animalis were the main users of the ~(13)C-labeled substrate. Culture-based studies of strains of these bacterial species enabled trophisms associated with inulin and its hydrolysis products to be identified. B. uniformis utilized Fibruline-inulin for growth, whereas the other species used fructo-oligosaccharide and monosaccharides. Thus, RNA–stable-isotope probing (RNA-SIP) provided new information about the use of carbon from inulin in microbiota metabolism.
机译:为了了解肠道​​菌群的复杂系统发育和功能,需要了解支持肠道菌群组成的营养。一次将稳定同位素(〜(13)C)标记的菊粉添加到大鼠的饮食中,以检测盲肠微生物群中特定成员对菊粉衍生底物的利用。在提供[〜(13)C]菊粉饮食后(0小时)之前和第6、12、18和24小时,收集来自Fibruline-菊粉喂养大鼠的盲肠消化物。从这些盲肠标本中提取RNA,并在等密度浮力密度梯度中进行分级分离,以检测〜(13)C标记的核酸,该核酸源自已代谢被标记饮食成分的细菌细胞。提供标记饮食后从标本中提取的RNA比0-h RNA密度更高。从这些级分获得的cDNA扩增的16S rRNA基因的测序表明,均匀的拟杆菌,葡糖杆菌,吲哚梭菌和动物双歧杆菌是〜(13)C标记底物的主要使用者。对这些细菌种类的菌株进行基于文化的研究,可以确定与菊粉及其水解产物相关的营养。统一芽孢杆菌利用纤维蛋白-菊粉促进生长,而其他物种则使用低聚果糖和单糖。因此,RNA稳定同位素探测(RNA-SIP)提供了有关菊粉在微生物群代谢中使用碳的新信息。

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