Diversity of anaerobic arsenite-oxidizing bacteria in low-salt environments analyzed with a newly developed PCR-based method
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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 the arxA gene. In this study, a novel primer pair for the arxA gene 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, diverse arxA gene 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 known arxA genes, found in the genome of alkaliphiles and halophiles.
KeywordsArsenic Arsenite oxidase arxA PCR detection Primer design
We are grateful to Arisa Shinohara for technical assistance. This work was supported by KAKENHI Grant Number 15K07209 to Kojima.
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Conflict of interest
The authors declare that there is no conflict of interest.
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