Document Type

Honors Project

First Advisor

Dr. Stephen Baron

Second Advisor

Dr. Gavin Lawson

Third Advisor

Dr. Edgar Lickey

Degree Award Date

Spring 2022

Keywords

Streptomyces, Promoter, Regulation of pHAZ gene, transformation, protoplast, gel electrophoresis, pcr, xyle gene, regulation, sequence, R5 media, reporter gene, Streptomyces sp. SFB5A

Disciplines

Biological Factors | Complex Mixtures | Enzymes and Coenzymes | Nucleic Acids, Nucleotides, and Nucleosides

Abstract

Streptomyces sp. SFB5A is an actinomycete (filamentous bacterium) that degrades PHB using an extracellular PHB depolymerase. PHB depolymerase synthesis is induced by growth on PHB and repressed by glucose (or other preferred carbon sources)2, which suggests that there is transcriptional regulation of its corresponding gene, phaZ. Binding sites are suspected to be the repeat sequences found in the phaZ gene. There are 3 different sets of primers that can be made in order to emphasize specific functional repeats while disrupting the functionality of others. The goal of the project is to transform Streptomyces sp. 5A with constructs containing a resistance gene and PCR products from the aforementioned primers, and then observe expression via culturing. The cells were attempted to be transformed via protoplast transformation. A more efficient method for protoplasting was discovered.

Recommended Citation

Baron, S., Senter, H. W. (2021). Discovering the role of repeat sequences in regulation of the phaZ gene in Streptomycetes sp. SFBSA. [Scholarly project]

Force Open Access

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