Presentation Type
Poster
Fluorescence In situ hybridization (FISH) of the tetrathionate reductase (ttr) gene in marine sulfur-reducing bacteria.
Abstract
Fluorescence in situ hybridization (FISH) has been used in numerous ways to identify the presence or absence of complementary nucleic acid sequences in a cell. Nucleotide probes are labeled with fluorescent molecules and designed to target genes or any complementary nucleic acid sequence of interest. In microbial ecology, FISH is used for the identification of microorganisms based on their 16s rRNA phylogeny, as well as for targeting specific functional genes on chromosomal DNA. This method will be applied to environmental cells in order to identify the presence and spatial distribution of the tetrathionate reductase gene (ttr), an anaerobic respiratory enzyme found in many species of sulfur reducing proteobacteria. The enzyme catalyzes the reductive cleavage of tetrathionate into two molecules of thiosulfate. Computer programs MEGA and BLAST were used to analyze and compare DNA sequences of the catalytic subunit, ttrA, from several different species. All conserved motifs were identified from the alignments, and probes were designed to target those areas using ARB. In silico experiments were performed using Sequencher to test the probes specificity. The qualifying probes will be used to identify the presence and distribution of the ttrA gene in a community of marine sulfur reducing bacteria from the Gulf.
Categories
Natural Sciences
Research Type
Research Assistant
Mentor Information
Haydn Rubelman
Fluorescence In situ hybridization (FISH) of the tetrathionate reductase (ttr) gene in marine sulfur-reducing bacteria.
Fluorescence in situ hybridization (FISH) has been used in numerous ways to identify the presence or absence of complementary nucleic acid sequences in a cell. Nucleotide probes are labeled with fluorescent molecules and designed to target genes or any complementary nucleic acid sequence of interest. In microbial ecology, FISH is used for the identification of microorganisms based on their 16s rRNA phylogeny, as well as for targeting specific functional genes on chromosomal DNA. This method will be applied to environmental cells in order to identify the presence and spatial distribution of the tetrathionate reductase gene (ttr), an anaerobic respiratory enzyme found in many species of sulfur reducing proteobacteria. The enzyme catalyzes the reductive cleavage of tetrathionate into two molecules of thiosulfate. Computer programs MEGA and BLAST were used to analyze and compare DNA sequences of the catalytic subunit, ttrA, from several different species. All conserved motifs were identified from the alignments, and probes were designed to target those areas using ARB. In silico experiments were performed using Sequencher to test the probes specificity. The qualifying probes will be used to identify the presence and distribution of the ttrA gene in a community of marine sulfur reducing bacteria from the Gulf.
