Identification and functional evaluation of a Temporin peptide, QUB-1549, from amphibian skin secretion
Amphibian species are a natural source for pharmaceuticals and their skin has been an area of research which has focused on skin secretions as a reservoir of active peptides. In the present study a new temporin peptide cDNA derived from the Hylarana latouchii was subject to “shotgun cloning”. The cDNAs were amplified using a polymerase chain reaction (PCR), and subject to agarose gel analysis. The products were purified and subject to cloning (ligation, transformation, isolation of recombinant plasmid DNA and PCR). Gel analysis was then performed, and the products purified, followed by initiation of a sequencing reaction. The extension products were then purified and sequenced. The project was concluded by a bioinformatics survey using an internet based the Basic Local Alignment Search Tool (BLAST) of the National Centre for Biotechnology Information (NCBI) that the cDNA precursor is belonging to temporin family, and the nucleotide sequence was deduced by alignment between this peptide and temporin-Ltb. The corresponding synthetic replicate, QUB-1549, was found to possess potent antibacterial activity against Gram positive bacteria, and fungi at micro molar concentration (with MIC=2 µM), but weakly against Gram negative bacteria (with MIC=16 µM). Also, at the concentration of more than 10 micro molar, this peptide was able to inhibit the proliferation of three cancer cells, along with a haemolytic effect. Thesis embargoed until 31st October 2024
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