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The methanol dehydrogenase structural gene mxaF and its use as a functional gene probe for methanotrophs and methylotrophs.

机译:甲醇脱氢酶结构基因mxaF及其用作甲烷营养生物和甲基营养生物的功能基因探针。

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摘要

The methanol dehydrogenase gene mxaF, encoding the large subunit of the enzyme, was amplified from the DNA of a number of representative methanotrophs, methyletrophs, and environmental samples by PCR using primers designed from regions of conserved amino acid sequence identified by comparison of three known sequences of the large subunit of methanol dehydrogenase. The resulting 550-bp PCR products were cloned and sequenced. Analysis of the predicted amino acid sequences corresponding to these mxaF genes revealed strong sequence conservation. Of the 172 amino acid residues, 47% were conserved among all 22 sequences obtained in this study. Phylogenetic analysis of these MxaF sequences showed that those from type I and type II methanotrophs form two distinct clusters and are separate from MxaF sequences of other gram-negative methylotrophs. MxaF sequences retrieved by PCR from DNA isolated from a blanket bog peat core sample formed a distinct phylogenetic cluster within the MxaF sequences of type II methanotrophs and may originate from a novel group of acidophilic methanotrophs which have yet to be cultured from this environment.
机译:甲醇脱氢酶基因mxaF编码该酶的大亚基,通过使用从保守氨基酸序列区域设计的引物,通过PCR设计从多个代表性的甲烷营养生物,甲基营养生物和环境样品的DNA中扩增而来,这些氨基酸通过比较三个已知序列而确定甲醇脱氢酶大亚基的组成。克隆得到的550 bp PCR产物并测序。对与这些mxaF基因相对应的预测氨基酸序列的分析显示了强烈的序列保守性。在该研究中获得的所有22个序列中,在172个氨基酸残基中,有47%是保守的。这些MxaF序列的系统发育分析表明,来自I型和II型甲烷营养生物的序列形成两个不同的簇,并且与其他革兰氏阴性甲基营养菌的MxaF序列分离。通过PCR从分离自泥炭沼泽泥炭核心样品的DNA中检索到的MxaF序列在II型甲烷营养生物的MxaF序列内形成了独特的系统发生簇,并且可能源自尚未从该环境培养的一组新的嗜酸性甲烷营养生物。

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