Journal
ENZYME AND MICROBIAL TECHNOLOGY
Volume 85, Issue -, Pages 19-24Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2016.01.001
Keywords
Selenate reductase; Primer design; DMSO; Membrane biofilm reactor
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Funding
- National Natural Science Foundation of China [21107091, 21377109, 21577123]
- National High Technology Research and Development Program of China [2013AA06A205]
- Fundamental Research Funds for the Central Universities [2014FZA6008]
- Public Welfare Project of the Science and Technology Department of Zhejiang Province [2015C33016]
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We designed a primer set to target selenate reductase (SerA) for detecting selenate reducing bacteria (SeRB). Our serA gene-based PCR primer set has high specificity in that it and positively amplified some SeRB, but not denitrifying bacteria (DB). Phylogenetic analysis of serA clone sequences of environmental samples from selenate-reducing membrane biofilm reactor (MBfR) biofilms showed that these sequences were closely grouped and had high similarity to selenate reductase gene sequences from SeRB Thauera selenatis and DB Dechloromonas; however, they were distant to other genes from dimethylsulfoxide (DMSO) enzyme family. Constructing a standard curve targeting the serA gene, we found that the good linearity for the qPCR assay when applied it to quantify SeRB in MBfR biofilms, and the gene copies of SeRB correlated well to the selenate removal percentages. Our results demonstrated the feasibility of using the serA gene-based PCR primer set to detect and quantify SeRB in environmental samples. (C) 2016 Elsevier Inc. All rights reserved.
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