Date Written

Spring 4-1-2016

Document Type

Distinction Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

Jennifer Bennett

First Committee Member

David Robertson

Second Committee Member

Jeffrey Lehman

Keywords

Streptomyces, Developmental genes, Transposon mutagenesis, c-di-GMP, Electroporation, monooxygenase

Subject Categories

Bacteriology | Pathogenic Microbiology

Abstract

Streptomyces species are gram positive filamentous soil bacteria found worldwide. They are known for their ability to synthesize over two-thirds of the antibiotics in use today. The Streptomyces genome is the largest bacterial genome to be sequenced thus far and contains the largest known amount of genes within a single bacterium. Many of these genes are involved in the complex program of development that is characteristic of this bacterial genus. The gene SCAB27101 was deleted to introduce a new mutation in S. scabies using the REDIRECT protocol. A phenotypic comparison has been initiated for visual colony morphology, microscopic appearance, ability to make biofilms and ability to infect potatoes and other root crop vegetables of the wild type strain and the mutant candidates that have been created. It was hypothesized that the mutation will cause the spores to develop faster which will in turn make the mutant more pathogenic towards the root crop vegetables. Surprisingly, the mutant candidates exhibited an obvious delay in development. In a separate project, S. coelicolor transposon mutant candidates exhibiting potential developmental defects were also analyzed using colony morphology and phase-contrast microscopy. To identify the site of transposon insertion, chromosomal DNA was isolated, subjected to restriction enzyme digestion and subsequent ligation followed by inverse polymerase chain reaction (PCR) and sequencing. There are many additional developmental genes that are believed to exist in this pharmacologically important bacterial genus, and this research provides multiple genes of interest for further study.

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