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...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English English |
| Published: |
Elsevier
2009
|
| Subjects: | |
| 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/10356/3/Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria.pdf |
| _version_ | 1825719844825202688 |
|---|---|
| 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 |
| collection | UM |
| 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. |
| first_indexed | 2024-03-06T05:25:57Z |
| format | Article |
| id | um.eprints-10356 |
| institution | Universiti Malaya |
| language | English English |
| last_indexed | 2024-03-06T05:25:57Z |
| publishDate | 2009 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | um.eprints-103562020-10-16T00:21:54Z http://eprints.um.edu.my/10356/ Development of a PCR assay for the detection 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 text 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 text en http://eprints.um.edu.my/10356/3/Development_of_a_PCR_assay_for_the_detection_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 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 <https://doi.org/10.1016/j.ijhydene.2009.04.029>. https://doi.org/10.1016/j.ijhydene.2009.04.029 doi:10.1016/j.ijhydene.2009.04.029 |
| 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 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 |
| topic | Q Science (General) QH Natural history QR Microbiology R Medicine |
| 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/10356/3/Development_of_a_PCR_assay_for_the_detection_of_nifH_and_nifD_genes_in_indigenous_photosynthetic_bacteria.pdf |
| work_keys_str_mv | AT tanjeeweng developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT thongkwailin developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT arumugamnirmaladevi developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT cheahwailian developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT laiyihwei developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT chuakekheng developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT rahimrahaabdul developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria AT vikineswarysabaratnam developmentofapcrassayforthedetectionofnifhandnifdgenesinindigenousphotosyntheticbacteria |