Development of a PCR assay for the detectiion 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, Jee Weng, Thong, Kwai Lin, Arumugam, Nirmala Devi, Cheah, Wai Lian, Lai, Yih Wei, Chua, Kek Heng, Rahim, Raha Abdul, Vikineswary, Sabaratnam
Format: Article
Language:English
Published: Elsevier 2009
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Online Access:http://eprints.um.edu.my/10356/1/2009_Development_of_a_PCR_assay_for_the_detectiion_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf
http://eprints.um.edu.my/10356/
https://doi.org/10.1016/j.ijhydene.2009.04.029
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Institution: Universiti Malaya
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spelling my.um.eprints.103562018-10-15T03:30:37Z http://eprints.um.edu.my/10356/ Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria Tan, Jee Weng Thong, Kwai Lin Arumugam, Nirmala Devi Cheah, Wai Lian Lai, Yih Wei Chua, Kek Heng Rahim, Raha Abdul Vikineswary, Sabaratnam Q Science (General) QH Natural history QR Microbiology 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/10356/1/2009_Development_of_a_PCR_assay_for_the_detectiion_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf 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 detectiion 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 Q Science (General)
QH Natural history
QR Microbiology
R Medicine
spellingShingle Q Science (General)
QH Natural history
QR Microbiology
R Medicine
Tan, Jee Weng
Thong, Kwai Lin
Arumugam, Nirmala Devi
Cheah, Wai Lian
Lai, Yih Wei
Chua, Kek Heng
Rahim, Raha Abdul
Vikineswary, Sabaratnam
Development of a PCR assay for the detectiion 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, Jee Weng
Thong, Kwai Lin
Arumugam, Nirmala Devi
Cheah, Wai Lian
Lai, Yih Wei
Chua, Kek Heng
Rahim, Raha Abdul
Vikineswary, Sabaratnam
author_facet Tan, Jee Weng
Thong, Kwai Lin
Arumugam, Nirmala Devi
Cheah, Wai Lian
Lai, Yih Wei
Chua, Kek Heng
Rahim, Raha Abdul
Vikineswary, Sabaratnam
author_sort Tan, Jee Weng
title Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria
title_short Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria
title_full Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria
title_fullStr Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria
title_full_unstemmed Development of a PCR assay for the detectiion of nifH and nifD genes in indigenous photosynthetic bacteria
title_sort development of a pcr assay for the detectiion of nifh and nifd genes in indigenous photosynthetic bacteria
publisher Elsevier
publishDate 2009
url http://eprints.um.edu.my/10356/1/2009_Development_of_a_PCR_assay_for_the_detectiion_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria..pdf
http://eprints.um.edu.my/10356/
https://doi.org/10.1016/j.ijhydene.2009.04.029
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