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: Jee, Weng Tan, Kwai , Lin Thong, Arumugam, Nirmala Devi, Wai , Lian Cheah, Yih , Wei Lai, Kek, Heng Chua, Abdul Rahim, Raha, Sabaratnam, Vikineswary
Format: Article
Language:English
English
Published: Elsevier 2009
Online Access:http://psasir.upm.edu.my/id/eprint/13993/
http://psasir.upm.edu.my/id/eprint/13993/1/Development%20of%20a%20PCR%20assay%20for%20the%20detection%20of%20nifH%20and%20nifD%20genes%20in%20indigenous%20photosynthetic%20bacteria.pdf
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author Jee, Weng Tan
Kwai , Lin Thong
Arumugam, Nirmala Devi
Wai , Lian Cheah
Yih , Wei Lai
Kek, Heng Chua
Abdul Rahim, Raha
Sabaratnam, Vikineswary
author_facet Jee, Weng Tan
Kwai , Lin Thong
Arumugam, Nirmala Devi
Wai , Lian Cheah
Yih , Wei Lai
Kek, Heng Chua
Abdul Rahim, Raha
Sabaratnam, Vikineswary
author_sort Jee, Weng Tan
building UPM Institutional Repository
collection Online Access
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.
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English
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publisher Elsevier
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spelling upm-139932015-10-09T00:48:31Z http://psasir.upm.edu.my/id/eprint/13993/ Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria. Jee, Weng Tan Kwai , Lin Thong Arumugam, Nirmala Devi Wai , Lian Cheah Yih , Wei Lai Kek, Heng Chua Abdul Rahim, Raha Sabaratnam, Vikineswary 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 2009-09 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/13993/1/Development%20of%20a%20PCR%20assay%20for%20the%20detection%20of%20nifH%20and%20nifD%20genes%20in%20indigenous%20photosynthetic%20bacteria.pdf 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 10.1016/j.ijhydene.2009.04.029 English
spellingShingle Jee, Weng Tan
Kwai , Lin Thong
Arumugam, Nirmala Devi
Wai , Lian Cheah
Yih , Wei Lai
Kek, Heng Chua
Abdul Rahim, Raha
Sabaratnam, Vikineswary
Development of a PCR assay for the detection of nifH and nifD genes in indigenous photosynthetic bacteria.
title 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_short 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.
url http://psasir.upm.edu.my/id/eprint/13993/
http://psasir.upm.edu.my/id/eprint/13993/
http://psasir.upm.edu.my/id/eprint/13993/1/Development%20of%20a%20PCR%20assay%20for%20the%20detection%20of%20nifH%20and%20nifD%20genes%20in%20indigenous%20photosynthetic%20bacteria.pdf