Kentucky State University
Sequence Identification of Ribonucleotide Reductase from Exiguiobacterium acetylicum Strain SN
Institution
Kentucky State University
Faculty Advisor/ Mentor
Williard Mazhawidza; Narayanan Rajendran
Abstract
The mechanism to biosynthesize elements of the cell membrane is essential to cell survival especially for prokaryotic cells. Such mechanisms are catalyzed by biosynthetic enzymes. To identify such a biosynthetase enzyme, we employed polymerase chain reaction (PCR) using a set of synthetase primers to probe genomic DNA of a soil isolate Exiguiobacterium acetylicum strain SN. The amplified PCR products were profiled by agarose gel electrophoresis. Selected product was cloned in our microbiology lab and sequenced at the University of Louisville DNA Core facility. The free software, MEGA 4.1, was employed in searching NCBI linked protein database and the DNA sequence analysis, which putatively, identified our DNA amplicon as ribonucleotide reductase(RNR). Multiple DNA sequence alignments revealed the conserved amino acids which is specific to this group of enzymes. Based on the Cn3D software a three dimensional modeling was made, which revealed that our cloned sequence fits with the established model of other ribonucleotide reductase. Phylogenetic analysis is underway to match the bacteria based on the matching sequences of Exiguiobacterium acetylicum strain SN.
Sequence Identification of Ribonucleotide Reductase from Exiguiobacterium acetylicum Strain SN
The mechanism to biosynthesize elements of the cell membrane is essential to cell survival especially for prokaryotic cells. Such mechanisms are catalyzed by biosynthetic enzymes. To identify such a biosynthetase enzyme, we employed polymerase chain reaction (PCR) using a set of synthetase primers to probe genomic DNA of a soil isolate Exiguiobacterium acetylicum strain SN. The amplified PCR products were profiled by agarose gel electrophoresis. Selected product was cloned in our microbiology lab and sequenced at the University of Louisville DNA Core facility. The free software, MEGA 4.1, was employed in searching NCBI linked protein database and the DNA sequence analysis, which putatively, identified our DNA amplicon as ribonucleotide reductase(RNR). Multiple DNA sequence alignments revealed the conserved amino acids which is specific to this group of enzymes. Based on the Cn3D software a three dimensional modeling was made, which revealed that our cloned sequence fits with the established model of other ribonucleotide reductase. Phylogenetic analysis is underway to match the bacteria based on the matching sequences of Exiguiobacterium acetylicum strain SN.