Diversity of anaerobic arsenite-oxidizing bacteria in low-salt environments analyzed with a newly developed PCR-based method
Diversity of anaerobic arsenite-oxidizing bacteria in low-salt environments analyzed with a newly developed PCR-based method
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DOI：
10.1007/s10201-018-0539-1
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发表时间：
2018-02
期刊：
影响因子：
1.6
通讯作者：
Melody Cabrera Ospino;Hisaya Kojima;Tomohiro Watanabe;Tomoya Iwata;M. Fukui
中科院分区：
文献类型：
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作者：
Melody Cabrera Ospino;Hisaya Kojima;Tomohiro Watanabe;Tomoya Iwata;M. Fukui
厌氧亚砷酸盐氧化是砷循环中潜在的重要但最不为人所知的过程。厌氧亚砷酸盐氧化关键酶的催化亚基由arxA基因编码。在这项研究中，设计了一对新的针对thearxA基因的引物来检测这个著名基因的不同序列。通过在其5‘端增加额外的碱基，对所设计的引物进行了进一步的修饰。这一改进使得用TA克隆分析PCR产物成为可能，从而提供了更高的研究吞吐量。通过改进的引物对和TA克隆相结合的方法，从湖水、泉水和温泉微生物席中有效地获得了多样化的arxA基因序列。在低盐度和近中性pH的样品中检测到的序列在系统发育上与大多数已知的在嗜碱和嗜盐细菌基因组中发现的xA基因不同。
Anaerobic arsenite oxidation is potentially important but the least understood process in the arsenic cycle. The catalytic subunit of the key enzyme for anaerobic arsenite oxidation is encoded by thearxAgene. In this study, a novel primer pair for thearxAgene was designed to detect diverse sequences of this notable gene. Further modification of the designed primer was made by adding extra bases to its 5′- end. This modification made it possible to analyze the PCR products with TA cloning, which provides higher throughput of investigations. With the combination of modified primer pair and TA cloning, diversearxAgene sequences were effectively obtained from samples of lake water, spring water, and hot spring microbial mat. The sequences detected in the samples characterized by low salinity and nearly neutral pH were phylogenetically distinct from the majority of previously knownarxAgenes, found in the genome of alkaliphiles and halophiles.