Date of Award

Spring 4-19-2018

Document Type

Distinction Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

Dr. Jennifer Bennett

First Committee Member

Dr. Jennifer Bennett

Second Committee Member

Dr. Carrigan Hayes

Third Committee Member

Dr. Steffanie Burk

Keywords

Streptomyces, chromosome segregation, transposon, bacterial development, bacteria

Subject Categories

Molecular Biology | Molecular Genetics

Abstract

Streptomyces coelicolor is a soil bacterium that is a model for bacterial development. It is a filamentous, sporulating bacterium known to produce many medically utilized antibiotics. The goal of this research was to examine several developmental mutants and characterize novel genes of interest. Previously generated random transposon insertion mutants were analyzed using visual and microscopic phenotyping. Mutants of interest were further pursued and each transposon disruption site was identified by Inverse PCR and DNA sequencing. One of the novel genes is suspected to be involved in DNA segregation and codes for a putative membrane protein. Staining with propidium iodide was conducted to visualize DNA under confocal microscopy. Preliminary screening shows a significantly higher anucleation rate when compared to wild type parental strain. This is the highest known rate of anucleation in Streptomyces segregation mutants. A modified CRISPR-Cas9 system is being utilized to construct a deletion of the putative chromosome segregation gene. Upon isolation of the deletion mutant, genetic complementation will be completed to confirm that the phenotype is caused by the gene of interest.

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