Kentucky State University

Poster Title

Targeted Amplification of a Matching Non-Ribosomal Peptide Synthetase Gene in Soil Bacterium Arthrobacter Nicotianae Strain PR

Institution

Kentucky State University

Abstract

Large numbers of biologically significant compounds are synthesized non-ribosomally by microorganisms. Non-ribosomal peptide synthetase (NRPS) is involved in this process to produce such peptide products. In order to find out the occurrence of such non-ribosomal biosynthetic system in Arthrobacter nicotianae strain PR, we used a degenerated set of NRPS primers. Along with basic microbial techniques such as spread plate, streak plate, broth culture, and centrifugation, we also used molecular techniques such as DNA isolation, Gel electrophoresis, and polymerase chain reaction (PCR). The genomic DNA was isolated using DNA bactozol kit. It was amplified with NRPS primer using our PCR protocols that are standardized earlier. PCR amplicons of the target strain obtained by using a specific set of forward and reverse primers was checked in a 1 % electrophoresis gel. Along with the amplicon of a control 16S primer, the target amplicons were photographed using UVP documentation unit. This targeted amplification approach revealed that the tested primer set of NRPS against the genomic DNA of A. nicotianae is matching with our newly isolated soil bacterium. Further cloning and sequence analysis are underway.

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Targeted Amplification of a Matching Non-Ribosomal Peptide Synthetase Gene in Soil Bacterium Arthrobacter Nicotianae Strain PR

Large numbers of biologically significant compounds are synthesized non-ribosomally by microorganisms. Non-ribosomal peptide synthetase (NRPS) is involved in this process to produce such peptide products. In order to find out the occurrence of such non-ribosomal biosynthetic system in Arthrobacter nicotianae strain PR, we used a degenerated set of NRPS primers. Along with basic microbial techniques such as spread plate, streak plate, broth culture, and centrifugation, we also used molecular techniques such as DNA isolation, Gel electrophoresis, and polymerase chain reaction (PCR). The genomic DNA was isolated using DNA bactozol kit. It was amplified with NRPS primer using our PCR protocols that are standardized earlier. PCR amplicons of the target strain obtained by using a specific set of forward and reverse primers was checked in a 1 % electrophoresis gel. Along with the amplicon of a control 16S primer, the target amplicons were photographed using UVP documentation unit. This targeted amplification approach revealed that the tested primer set of NRPS against the genomic DNA of A. nicotianae is matching with our newly isolated soil bacterium. Further cloning and sequence analysis are underway.