Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria
Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria
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DOI：
10.1099/00221287-148-11-3651
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发表时间：
2002-11-01
期刊：
影响因子：
2.8
通讯作者：
Bos, NA
中科院分区：
文献类型：
--
作者：
Salzman, NH;de Jong, H;Bos, NA
以完整的小鼠小肠、大肠、盲肠和粪便标本的总基因组DNA为模板，用保守的细菌引物进行16S rRNA基因序列的PCR扩增。对扩增产物进行了系统发育分析，发现了40个独特的16S rDNA序列。在这些序列中，25%(10/40)与所描述的小鼠肠道微生物相对应，包括乳杆菌、幽门螺杆菌、节段丝状细菌和改变的Schaedler菌群成员(ASF360、ASF361、ASF502和ASF519)；75%(30/40)代表新序列。大量的新序列(11/40)揭示了一个新的可操作的分类单位(OTU)，属于细胞吞噬-黄杆菌-类杆菌门，作者将其命名为‘小鼠肠道细菌’。16S rRNA探针是为这种新的OTU而研制的。通过对新序列的分析，发现有8个聚在直肠杆菌-球状梭菌组内，3个聚在类杆菌组内。其中一个新序列与刺毛韦氏杆菌有较远的亲缘关系，另一个与霉菌芽孢杆菌有较远的亲缘关系。产生了针对这些新克隆的165rRNA的寡核苷酸探针。结合使用之前描述的四个探针和新设计的四个探针，从小鼠大肠中回收的大约80%的细菌和从小鼠盲肠中回收的71%的细菌可以通过荧光原位杂交(FISH)进行鉴定。
Total genomic DNA from samples of intact mouse small intestine, large intestine, caecum and faeces was used as template for PCR amplification of 16S rRNA gene sequences with conserved bacterial primers. Phylogenetic analysis of the amplification products revealed 40 unique 16S rDNA sequences. Of these sequences, 25% (10/40) corresponded to described intestinal organisms of the mouse, including Lactobacillus spp., Helicobacter spp., segmented filamentous bacteria and members of the altered Schaedler flora (ASF360, ASF361, ASF502 and ASF519); 75% (30/40) represented novel sequences. A large number (11/40) of the novel sequences revealed a new operational taxonomic unit (OTU) belonging to the Cytophaga-Flavobacter-Bacteroides phylum, which the authors named 'mouse intestinal bacteria'. 16S rRNA probes were developed for this new OTU. Upon analysis of the novel sequences, eight were found to cluster within the Eubacterium rectale-Clostridium coccoides group and three clustered within the Bacteroides group. One of the novel sequences was distantly related to Verrucomicrobium spinosum and one was distantly related to Bacillus mycoides. Oligonucleotide probes specific for the 165 rRNA of these novel clones were generated. Using a combination of four previously described and four newly designed probes, approximately 80% of bacteria recovered from the murine large intestine and 71% of bacteria recovered from the murine caecum could be identified by fluorescence in situ hybridization (FISH).