Kentucky State University

Poster Title

Molecular Probing Of Soil Bacterium Exiguobacterium Acetylicum Strain SN Using Degenerated Primers

Institution

Kentucky State University

Abstract

Many soil bacteria and filamentous fungi produce some valuable peptides such as cyclosporine, penicillin etc non-ribosomally. A multienzyme complex, known as nonribosomal peptide synthetase (NRPS), is involved in this process. The objective of this study was to find out the presence of such non-ribosomal system from newly isolated soil bacterium Exiguobacterium acetylicum strain SN and to get hands-on training in various molecular and microbiology techniques used in this research. As expected, this study could also give us a clue for the presence of a peptide product such as an antibiotic in this soil bacterium. In order to achieve these objectives, initially a pure Nutrient Broth culture of E. acetylicum was grown at 30° C, by picking a single colony from a Nutrient Agar culture plate prepared earlier by streak-plate technique. Genomic DNA was isolated using DNA bactozol kit and confirmed with 1 % agarose gel. Different sets degenerated probes of NRPS were used to amplify the DNA using Polymerase Chain Reaction (PCR) technique. The employed PCR reactions were depended upon the PCR cycles we standardized earlier. The PCR amplicons of the genomic DNA were studied using similar electrophoresis method. By using UVP documentation unit, we examined the targeted amplicons with control primer products. Our results confirmed the presence of non-ribosomal system in E. acetylicum as proved by the targeted-primer set against the genomic DNA along with the control of 16S primer. Further analysis, cloning of the amplicon and the DNA sequence studies are underway.

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Molecular Probing Of Soil Bacterium Exiguobacterium Acetylicum Strain SN Using Degenerated Primers

Many soil bacteria and filamentous fungi produce some valuable peptides such as cyclosporine, penicillin etc non-ribosomally. A multienzyme complex, known as nonribosomal peptide synthetase (NRPS), is involved in this process. The objective of this study was to find out the presence of such non-ribosomal system from newly isolated soil bacterium Exiguobacterium acetylicum strain SN and to get hands-on training in various molecular and microbiology techniques used in this research. As expected, this study could also give us a clue for the presence of a peptide product such as an antibiotic in this soil bacterium. In order to achieve these objectives, initially a pure Nutrient Broth culture of E. acetylicum was grown at 30° C, by picking a single colony from a Nutrient Agar culture plate prepared earlier by streak-plate technique. Genomic DNA was isolated using DNA bactozol kit and confirmed with 1 % agarose gel. Different sets degenerated probes of NRPS were used to amplify the DNA using Polymerase Chain Reaction (PCR) technique. The employed PCR reactions were depended upon the PCR cycles we standardized earlier. The PCR amplicons of the genomic DNA were studied using similar electrophoresis method. By using UVP documentation unit, we examined the targeted amplicons with control primer products. Our results confirmed the presence of non-ribosomal system in E. acetylicum as proved by the targeted-primer set against the genomic DNA along with the control of 16S primer. Further analysis, cloning of the amplicon and the DNA sequence studies are underway.