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

Tan, J.W.; Thong, K.L.; Arumugam, N.D.; Cheah, W.L.; Lai, Y.W.; Chua, K.H.; Rahim, R.A.; Vikineswary, S. (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

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    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.

    Item Type: Article
    Creators:
    1. Tan, J.W.
    2. Thong, K.L.(Institute of Biological Sciences, Faculty of Science Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
    3. Arumugam, N.D.
    4. Cheah, W.L.
    5. Lai, Y.W.
    6. Chua, K.H.(Department of Molecular Medicine, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
    7. Rahim, R.A.
    8. Vikineswary, S.
    Journal or Publication Title: International Journal of Hydrogen Energy
    Additional Information: Department of Molecular Medicine, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA
    Uncontrolled Keywords: Biohydrogen; Nitrogenase; PCR detection; nifH; nifD; Photosynthetic bacteria
    Subjects: R Medicine
    Divisions: Faculty of Medicine
    Depositing User: Ms azrahani halim
    Date Deposited: 01 Nov 2012 15:52
    Last Modified: 15 Dec 2014 14:29
    URI: http://eprints.um.edu.my/id/eprint/3804

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