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免培养法研究野生川金丝猴肠道内生细菌多样性
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陕西省科技厅科技统筹创新计划项目(No. 2012HBGC-20);陕西理工学院人才引进启动项目


Culture-independent analysis of intestinal microbial diversity from Rhinopithecus roxellana
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    摘要:

    【目的】了解野生川金丝猴(Rhinopithecus roxellana)肠道内生细菌的组成及其多样性。【方法】提取川金丝猴肠道内生细菌总DNA,选用细菌通用引物799F和1492R对总DNA进行16S rRNA基因特异性扩增,构建川金丝猴肠道内生细菌16S rRNA基因克隆文库,对阳性克隆进行限制性内切酶片段长度多态性(PCR-RFLP)分析,并对Hae Ⅲ酶切带谱菌株进行测序,构建系统发育树。【结果】根据酶切带谱分析和测序结果,将随机挑取的157个阳性克隆归为27个不同的可操作分类单元(OTUs)。系统发育分析表明这些克隆序列有62.10%属于厚壁菌门(Firmicutes),其中包括梭菌属(Clostridium)、Cellulosilyticum属、Robinsoniella属、Anaerofustis属、Blautia属和Anaerovorax属,有37.90%属于未培养细菌。【结论】川金丝猴肠道内生细菌多样性丰富,并且可能存在新的分类单元。

    Abstract:

    [Objective] The composition and diversity of intestinal microbial diversity of wild Sichuan snub-nosed monkey (Rhinopithecus roxellana) were investigated. [Methods] The total DNA extracted from endophytic bacterial was performed 16S rRNA gene specific amplification by used bacteria-specific primers 799F and 1492R and constructed a clone library. Then, 16S rRNA gene phylogenetic tree was established by analyzed positive clones used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and by sequenced some of the clones. [Results] The 157 clones analyzed were grouped into 27 operational taxonomic units (OTUs), where the clone distribution was as follows: 62.10% of Firmicutes which contained Clostridium, Cellulosilyticum, Robinsoniella, Anaerofustis, Anaerovorax and Blautia, 37.90% of uncultured bacteria. [Conclusion] The intestine bacterial of wild Sichuan snub-nosed monkey has a rich diversity and maybe exist new taxon.

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杨曼,兰阿峰,郭素芬,丁小维,邓百万. 免培养法研究野生川金丝猴肠道内生细菌多样性[J]. 微生物学通报, 2014, 41(8): 1605-1612

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  • 在线发布日期: 2014-08-07
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