Using new polymerase chain reaction (PCR) primers, a once known to be under-transcribed microcystin synthetase A (mcyA) gene from the only known toxigenic cyanobacterium Microcystis aeruginosa dominating the Hartbeespoort Dam was consistently amplified from genomic DNA extracted from a set of algal and cell free water samples collected across this dam. In addition to this, five more mcy genes (mcyBCDEG) were also amplified during this study. The resultant mcyA PCR products (518 bp) were purified and sequenced and gave nucleotide sequence segments of 408 bp sizes. The obtained sequence was aligned to the published mcyA gene sequence available online on the NCBI database and resulted in 100% similarity to a 408 bp mcyA gene sequence segment of M. aeruginosa UWOCC RID-1. Furthermore, it was found that the above sequence segment (408 bp) spans from a common base in M. aeruginosa PCC 7806 and M. aeruginosa PCC 7820 from 141 to 548 bp in the N-methyl transferase (NMT) region signifying their closer relatedness to M. aeruginosa UWOCC strains. This study has for the first time amplified mcyA gene consistently from both intracellular and extracellular DNA extracts obtained from algal and cell free water samples, respectively. Sequence data and the amplified mcy genes showed that M. aeruginosa is widely distributed and dominant in this dam.
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Research Article|
April 25 2013
PCR amplification and DNA sequence of mcyA gene: The distribution profile of a toxigenic Microcystis aeruginosa in the Hartbeespoort Dam, South Africa
Elbert A. Mbukwa;
Elbert A. Mbukwa
1Department of Applied Chemistry, Faculty of Science, University of Johannesburg, P.O. Box 17011, Doornfontein 2028, Johannesburg, Republic of South Africa
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Sammy Boussiba;
Sammy Boussiba
2Microalgal Biotechnology Laboratory, Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, Sede-Boker Campus, 84990, Israel
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Victor Wepener;
Victor Wepener
3School of Biological Sciences, Potchefstroom Campus, North West University, Private Bag X6001, Potchefstroom 2520, Republic of South Africa
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Stefan Leu;
Stefan Leu
2Microalgal Biotechnology Laboratory, Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, Sede-Boker Campus, 84990, Israel
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Yuval Kaye;
Yuval Kaye
2Microalgal Biotechnology Laboratory, Jacob Blaustein Institutes for Desert Research, Ben-Gurion University of the Negev, Sede-Boker Campus, 84990, Israel
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Titus A. M. Msagati;
1Department of Applied Chemistry, Faculty of Science, University of Johannesburg, P.O. Box 17011, Doornfontein 2028, Johannesburg, Republic of South Africa
E-mail: [email protected]
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Bhekie B. Mamba
Bhekie B. Mamba
1Department of Applied Chemistry, Faculty of Science, University of Johannesburg, P.O. Box 17011, Doornfontein 2028, Johannesburg, Republic of South Africa
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J Water Health (2013) 11 (3): 563–572.
Article history
Received:
January 03 2013
Accepted:
April 01 2013
Citation
Elbert A. Mbukwa, Sammy Boussiba, Victor Wepener, Stefan Leu, Yuval Kaye, Titus A. M. Msagati, Bhekie B. Mamba; PCR amplification and DNA sequence of mcyA gene: The distribution profile of a toxigenic Microcystis aeruginosa in the Hartbeespoort Dam, South Africa. J Water Health 1 September 2013; 11 (3): 563–572. doi: https://doi.org/10.2166/wh.2013.014
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