Document Type

Honors Project

First Advisor

Dr. Stephen F. Baron

Second Advisor

Dr. Edgar Lickey

Third Advisor

Dr. Timothy A. Kreps

Degree Award Date

Spring 2022

Keywords

Electroporation, Transformation, Streptomyces nymphaeiformis, Stationary phase, Mid-log phase, Spores, Flocculation

Disciplines

Biology | Biotechnology | Cell Biology | Environmental Microbiology and Microbial Ecology

Abstract

Streptomyces species are notoriously difficult to transform. Streptomyces nymphaeiformis is no different, so a method of electroporation was used to attempt to transform the cells. Multiple growth stages were used in order to alter the degree of development of the cell wall. The procedure did not kill the cells, but the cells were not transformed. Due to the lack of transformation with S. nymphaeiformis, transformation was attempted on two other Streptomyces strains, S. lividans and S. coelicolor. Neither was successfully transformed to thiostrepton (tsr) resistance, nor did they grow on a plate lacking thiostrepton. One possibility for the lack of transformation and growth was flocculation of the cells in the broth after electroporation. The flocculation may have prevented the electric pulse from shocking all the cells or transferring the sample to the plate for selection was made more difficult. Finding a method to prevent flocculation may allow for successful transformation using an electroporation method.

Recommended Citation

Knott, Heather, & Baron, Stephen. "Developing an Electroporation Method for Transforming Streptomyces nymphaeiformis." Honors Projects, Bridgewater College, 2022.

Force Open Access

1

Knott Heather Electroporation ASPIRE 040122.pdf (619 kB)
Developing an Electroporation Method for Transforming Streptomyces nymphaeiformis Poster

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