Browse all Channels
IWA Publishing
Alliance House, 12 Caxton Street ,
London ,
United Kingdom SW1H 0QS
United Kingdom
  4. PCR amplification and DNA sequence of mcyA gene: The ...

PCR amplification and DNA sequence of mcyA gene: The distribution profile of a toxigenic Microcystis aeruginosa in the Hartbeespoort Dam, South Africa

0
- By: , , , , , ,

Courtesy of IWA Publishing

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.

Related articles

  • qPCR detection versus microscopy observations for assessing presence–absence of Didymosphenia ...

    Microscopic versus qPCR (quantitative polymerase chain reaction) analyses were compared for the detection of Didymosphenia geminata in biofilms and water filtrates from seven Gaspésie rivers (Canada). For the qPCR approach, two DNA extraction kits (QIAamp DNA Micro Kit, Qiagen and PowerSoil DNA Isolation Kit, Mo Bio Laboratories) and two pairs of primers were considered. The pair of primers D602F/D753Rext did not amplify D. geminata DNA whereas the pair of primers D602F/D753R was specific for D. geminata. Presence-...

  • Ecology of duckweed ponds used for nutrient recovery from wastewater

    The microorganism community that grows under duckweed shelter can play an important role on treatment processes. Therefore, the present study aimed to assess the zooplankton dynamic and microbial community in duckweed ponds (DPs) applied for domestic wastewater treatment under open field conditions. A pilot system comprised of two DPs in series (DP1 and DP2), with 10 m2 each, received domestic wastewater through a flow rate of 200 L·day−1. Thus, the system was monitored during 314 days through samples collected...

  • ELITe InGenius a new Molecular Diagnostics Sample-to-Result solution and first fully automated ...

    ELITe InGenius is an integrated sample to-result solution dedicated to Molecular Diagnostics which automatically performs extraction, amplification and result interpretation with an unprecedented flexibility and assay menu possibility. The system combines a universal extraction process in unitary cartridge with individual and independently controlled RT PCR in cassette based format enabling the laboratories to develop and run custom panels of assays on demand. ELITe InGenius is an innovative solution build on our...

  • A new immunomagnetic bead separation–surfactant extraction treatment protocol for rapid and ...

    The Cryptosporidium oocyst is encased in a robust wall that is extremely resistant to detrimental environmental factors such as chlorine used to disinfect potable water. Therefore, extracting oocyst DNA is not a trivial undertaking. Standard procedures used to extract DNA from oocysts, such as freeze–thaw (F/T) methods and DNA purification kits, are time-consuming and expensive and are difficult to implement in routine clinical practice. Therefore, we developed a surfactant extraction treatment (SET) that efficient...

  • The DNA Revolution for Water Professionals

    Imagine trying to solve a problem when you are provided just 1% of the total data. This is essentially the case in analyzing water and wastewater microbiomes. Conventional microbiological testing methods such as culture and microscopy can identify only about 1% of microorganisms that are contained in a typical environmental sample. The remaining 99% are known to be there, but can not be identified by traditional culture tests. This 99% of unidentifiable microbes are sometimes referred to as “microbial dark...

Customer comments

No comments were found for PCR amplification and DNA sequence of mcyA gene: The distribution profile of a toxigenic Microcystis aeruginosa in the Hartbeespoort Dam, South Africa. Be the first to comment!