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Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria.


Citation

Jee, Weng Tan and Kwai , Lin Thong and Arumugam, Nirmala Devi and Wai , Lian Cheah and Yih , Wei Lai and Kek, Heng Chua and Abdul Rahim, Raha and Sabaratnam, Vikineswary (2009) Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria. International Journal of Hydrogen Energy, 34 (17). pp. 7538-7541. ISSN 0360-3199

Abstract

Molybdenum (Mo) nitrogenases consist of two components: dinitrogenase reductase (encoded by nifH) and the dinitrogenase or MoFe protein (encoded by nifDK). Nitrogenase enzyme of photosynthetic bacteria is responsible for hydrogen production. Therefore, primers were designed for the nitrogenase gene only. In this study, two primers (ND and NH) were designed after comparative genomic analysis of nifH and nifD gene sequences from public databases. The designed primers were used for the amplification of nifH and nifD genes to detect nitrogenase genes in photosynthetic bacteria. Initial detection was done using a monoplex Polymerase Chain Reactions (PCRs) followed by optimization of the PCR protocols. Subsequently, a duplex PCR was designed for amplification and detection of nifH and nifD genes in indigenous photosynthetic bacteria. Evaluation of the duplex PCR on six samples isolated from Palm Oil Mill Effluent (POME) showed that only four isolates contained both the nifH and nifD genes, indicating that these isolates were potential hydrogen-producing bacteria. PCR detection provides a rapid and efficient pre-identification of potential photosynthetic bacterial hydrogen producers.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Science
DOI Number: https://doi.org/10.1016/j.ijhydene.2009.04.029
Publisher: Elsevier
Keywords: Biohydrogen; Nitrogenase; PCR detection; nifH; nifD; Photosynthetic bacteria.
Depositing User: Nida Hidayati Ghazali
Date Deposited: 22 Jul 2013 04:43
Last Modified: 09 Oct 2015 00:48
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.ijhydene.2009.04.029
URI: http://psasir.upm.edu.my/id/eprint/13993
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