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通过古菌特异性聚合酶链反应和变性梯度凝胶电泳对产甲烷菌进行分析时,对古菌16S rRNA基因不同高变区的评估。

Evaluations of different hypervariable regions of archaeal 16S rRNA genes in profiling of methanogens by Archaea-specific PCR and denaturing gradient gel electrophoresis.

作者信息

Yu Zhongtang, García-González Rubén, Schanbacher Floyd L, Morrison Mark

机构信息

The MAPLE Research Initiative, Department of Animal Sciences, The Ohio State University, Columbus, Ohio 43210, USA.

出版信息

Appl Environ Microbiol. 2008 Feb;74(3):889-93. doi: 10.1128/AEM.00684-07. Epub 2007 Dec 14.

Abstract

Different hypervariable (V) regions of the archaeal 16S rRNA gene (rrs) were compared systematically to establish a preferred V region(s) for use in Archaea-specific PCR-denaturing gradient gel electrophoresis (DGGE). The PCR products of the V3 region produced the most informative DGGE profiles and permitted identification of common methanogens from rumen samples from sheep. This study also showed that different methanogens might be detected when different V regions are targeted by PCR-DGGE. Dietary fat appeared to transiently stimulate Methanosphaera stadtmanae but inhibit Methanobrevibacter sp. strain AbM4 in rumen samples.

摘要

对古菌16S rRNA基因(rrs)的不同高变(V)区域进行了系统比较,以确定用于古菌特异性PCR-变性梯度凝胶电泳(DGGE)的首选V区域。V3区域的PCR产物产生了最具信息性的DGGE图谱,并能够从绵羊瘤胃样本中鉴定出常见的产甲烷菌。该研究还表明,当PCR-DGGE针对不同的V区域时,可能会检测到不同的产甲烷菌。日粮脂肪似乎会短暂刺激史氏甲烷短杆菌,但会抑制瘤胃样本中的瘤胃甲烷短杆菌AbM4菌株。

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