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

Tan, Jee Weng and Thong, Kwai Lin and Arumugam, Nirmala Devi and Cheah, Wai Lian and Lai, Yih Wei and Chua, Kek Heng and Rahim, Raha Abdul and Vikineswary, Sabaratnam (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, DOI https://doi.org/10.1016/j.ijhydene.2009.04.029.

	Text 2009_Development_of_a_PCR_assay_for_the_detectiion_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf - Accepted Version Restricted to Repository staff only Download (427kB) | Request a copy
	Text Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria.pdf - Published Version Restricted to Repository staff only Download (181kB) | Request a copy

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
Funders:	UNSPECIFIED
Uncontrolled Keywords:	Biohydrogen; Nitrogenase; PCR detection; nifH; nifD; Photosynthetic bacteria
Subjects:	Q Science > Q Science (General) Q Science > QH Natural history Q Science > QR Microbiology R Medicine
Divisions:	Faculty of Science > Institute of Biological Sciences
Depositing User:	Prof. Dr Kwai Lin Thong
Date Deposited:	23 Oct 2014 04:34
Last Modified:	16 Oct 2020 00:21
URI:	http://eprints.um.edu.my/id/eprint/10356

Actions (login required)

View Item