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

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...

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Main Authors: Tan, J.W., Thong, Kwai Lin, Arumugam, N.D., Cheah, W.L., Lai, Y.W., Chua, Kek Heng, Rahim, R.A., Vikineswary, S.
Format: Article
Language:English
Published: Elsevier 2009
Subjects:
R Medicine
Online Access:http://eprints.um.edu.my/3804/1/Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf
http://eprints.um.edu.my/3804/
https://doi.org/10.1016/j.ijhydene.2009.04.029
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spelling my.um.eprints.38042018-10-15T03:32:08Z http://eprints.um.edu.my/3804/ Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria Tan, J.W. Thong, Kwai Lin Arumugam, N.D. Cheah, W.L. Lai, Y.W. Chua, Kek Heng Rahim, R.A. Vikineswary, S. R Medicine 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. Elsevier 2009 Article PeerReviewed application/pdf en http://eprints.um.edu.my/3804/1/Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf Tan, J.W. and Thong, Kwai Lin and Arumugam, N.D. and Cheah, W.L. and Lai, Y.W. and Chua, Kek Heng and Rahim, R.A. and 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 https://doi.org/10.1016/j.ijhydene.2009.04.029 doi:10.1016/j.ijhydene.2009.04.029
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
language English
topic R Medicine
spellingShingle R Medicine
Tan, J.W.
Thong, Kwai Lin
Arumugam, N.D.
Cheah, W.L.
Lai, Y.W.
Chua, Kek Heng
Rahim, R.A.
Vikineswary, S.
Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
description 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.
format Article
author Tan, J.W.
Thong, Kwai Lin
Arumugam, N.D.
Cheah, W.L.
Lai, Y.W.
Chua, Kek Heng
Rahim, R.A.
Vikineswary, S.
author_facet Tan, J.W.
Thong, Kwai Lin
Arumugam, N.D.
Cheah, W.L.
Lai, Y.W.
Chua, Kek Heng
Rahim, R.A.
Vikineswary, S.
author_sort Tan, J.W.
title Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
title_short Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
title_full Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
title_fullStr Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
title_full_unstemmed Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria
title_sort development of a pcr assay for the detection of nifh and nifd genes in indigenous photosynthetic bacteria
publisher Elsevier
publishDate 2009
url http://eprints.um.edu.my/3804/1/Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf
http://eprints.um.edu.my/3804/
https://doi.org/10.1016/j.ijhydene.2009.04.029
_version_ 1643687202759114752
score 13.211869

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