title: PCR amplification and DNA sequence of mcyA gene: the distribution profile of a toxigenic Microcystis aeruginosa in the Hartbeespoort Dam, South Africa.
abstract: 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.
authors: Department of Applied Chemistry, University of Johannesburg, Johannesburg, Republic of South Africa.
authors: Elbert A
authors: EA
authors: Mbukwa
authors: Sammy
authors: S
authors: Boussiba
authors: Victor
authors: V
authors: Wepener
authors: Stefan
authors: S
authors: Leu
authors: Yuval
authors: Y
authors: Kaye
authors: Titus A M
authors: TA
authors: Msagati
authors: Bhekie B
authors: BB
authors: Mamba
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005618:Fresh Water / Q000382:microbiology*; D005798:Genes, Bacterial; D046931:Microcystis / Q000235:genetics*; D010453:Peptide Synthases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013019:South Africa; D014871:Water Microbiology
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; D010453:Peptide Synthases; C432283:microcystin synthetase A