Location
University of Nevada Las Vegas, Student Union Ball Room
Start Date
6-8-2008 9:00 AM
End Date
6-8-2008 12:00 PM
Description
The goal of our research is to determine whether the level of transcription of a gene is correlated with the level of mutation in that gene. One factor involved in the mutability of a transcribed gene is the ability of the single stranded DNA to form secondary stem loop structures (SLS), in the wake of the transcription bubble, that contain unpaired mutable bases. We are interested in correlating the levels of mutation with transcription in the thiF gene, which is predicted by bioinformatic analysis to be highly mutable. To achieve this goal, Kathleen will first construct a non-polar thiF genetic knockout using a chloramphenicol cassette. Then, she will test the phenotype of the ThiF- strain. She will also build an IPTG-inducible construct containing thiF with a stop codon in the loop of a putative SLS. This will be introduced into ThiF- Bacillus subtilis and assayed for the accumulation of Thy+ mutations under starvation conditions, in the presence and absence of IPTG.
Keywords
Bacillus subtilis; Gene mutations; Isopropyl β-D-1-thiogalactopyranoside (IPTG); Mutagenesis; Secondary stem loop structures (SLS); ThiF
Disciplines
Genetics | Molecular Genetics
Language
English
Construction of a thiF genetic disruption in Bacillus subtilis
University of Nevada Las Vegas, Student Union Ball Room
The goal of our research is to determine whether the level of transcription of a gene is correlated with the level of mutation in that gene. One factor involved in the mutability of a transcribed gene is the ability of the single stranded DNA to form secondary stem loop structures (SLS), in the wake of the transcription bubble, that contain unpaired mutable bases. We are interested in correlating the levels of mutation with transcription in the thiF gene, which is predicted by bioinformatic analysis to be highly mutable. To achieve this goal, Kathleen will first construct a non-polar thiF genetic knockout using a chloramphenicol cassette. Then, she will test the phenotype of the ThiF- strain. She will also build an IPTG-inducible construct containing thiF with a stop codon in the loop of a putative SLS. This will be introduced into ThiF- Bacillus subtilis and assayed for the accumulation of Thy+ mutations under starvation conditions, in the presence and absence of IPTG.
Comments
Abstract & poster