Project Director

Harris, Bradley Jordan

Department Examiner

Turgeson, Andrew

Department

Dept. of Civil and Chemical Engineering

Publisher

University of Tennessee at Chattanooga

Place of Publication

Chattanooga (Tenn.)

Abstract

Genetic transformation of bacteria is a foundational technique in molecular biology and biotechnology, enabling the introduction and expression of foreign genes for research, industrial, and educational applications. This thesis investigates the transformation efficiency and green fluorescent protein (GFP) expression in Escherichia coli using the pGLO plasmid system, with the primary objective of supporting the development of a graduate-level laboratory course focused on bioprocess engineering and molecular biotechnology. The pGLO plasmid contains a GFP reporter gene regulated by an arabinose-inducible promoter and an ampicillin resistance gene that allows for selective growth of transformed cells. Competent E. coli HB101 K-12 cells were transformed using a calcium chloride-mediated heat shock protocol and subsequently cultured on selective media with and without arabinose. Successful transformation was confirmed qualitatively through bacterial growth on ampicillin-containing agar plates and visualization of GFP expression under ultraviolet illumination. Fluorescent colonies were consistently observed under inducing conditions, indicating effective plasmid uptake and regulated gene expression. Control plates lacking plasmid DNA or arabinose demonstrated expected growth patterns, supporting the validity of the experimental design. Although extensive quantitative measurements of transformation efficiency were limited, the experimental outcomes aligned with established behavior reported in the literature and instructional laboratory standards. This study emphasizes qualitative assessment of transformation success, experimental reproducibility, and biological variability inherent to bacterial gene expression systems. The results demonstrate that the pGLO transformation system functions reliably as a model for future integration into advanced bioprocess and bioengineering curricula.

Acknowledgments

I would like to sincerely thank Dr. Bradley Harris, Department Head, and Dr. Turgeson, Department Examiner, for their guidance, support, and advisement throughout this thesis process. Their feedback, encouragement, and academic insight were instrumental in the development and completion of this research. I would also like to extend my appreciation to my classmate and peer, Katie Collins, for her assistance and support during my research. Her help and encouragement were greatly valued and contributed to the success of this project. I am thankful for the opportunity to complete this thesis on transformation efficiency and GFP expression in E. coli, as it has been an important and rewarding part of my academic journey.

Degree

B. S.; An honors thesis submitted to the faculty of the University of Tennessee at Chattanooga in partial fulfillment of the requirements of the degree of Bachelor of Science.

Date

5-2026

Subject

Escherichia coli--Genetics; Genetic transformation; Green fluorescent proteins

Keyword

Transformation efficiency; GFP expression; E. coli; bacterial transformation; pGLO system; colony morphology

Discipline

Biochemical and Biomolecular Engineering

Document Type

Theses

Extent

iii, 25 leaves

DCMI Type

Text

Language

English

Rights

http://rightsstatements.org/vocab/InC/1.0/

License

http://creativecommons.org/licenses/by/4.0/

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