DOI
https://doi.org/10.25772/FXKB-C183
Defense Date
2004
Document Type
Dissertation
Degree Name
Doctor of Philosophy
Department
Microbiology & Immunology
First Advisor
Fang-Sheng Wu
Second Advisor
Guy Cabral
Third Advisor
Kathleen McCoy
Fourth Advisor
Jennifer Stewart
Fifth Advisor
L. Shozo Ozaki
Abstract
This dissertation describes a study utilizing bioinformatics to analyze homologues of a molecular chaperone, glucose-regulated protein 78 (grp 78), also known as BiP. The selected homologous proteins originate from organisms of infinitely diverse genera. Comparisons of protein sequence yielded the first clues of a common ancestry among these proteins. Furthermore, protein molecular weights, isoelectric points, N-terminal amino acids and half-lives of a known homolog and a non-homologous protein were examined. Additionally, electroporation, a state-of-the-art plasmid insertion technique, was explored using Chlamydomonas reinhardtii, a green alga, as the recipient of a parent plasmid, pSP124S. Distinctive hypertonic solutions and three separate field strengths were used in the plasmolysis of the cell wall of C. reinhardtii and subsequent electroporation, respectively. The number of transformants was tallied to evaluate which electroporation condition would yield the most transformed colonies. We had two discrete hypotheses: 1) that a structurally and functionally similar protein to glucose-regulated protein 78 exists across a wide spectrum of organisms and 2) that Chlamydomonas reinhardtii could be successfully transformed with pSP124S under certain electroporation conditions. The bioinformatics investigation revealed that analogous proteins to Human GRP 78 existed in Mus musculus (mouse), Rattus norvegicus (rat), Gallus domesticus (chicken), Gallus domesticus (chicken), Mesocricetus auratus (golden hamster), Bos taurus (cow), Xenopus laevis (frog), and Spinacia oleracea (spinach). Moreover, these homologous proteins more likely have a common evolutionary origin.
Rights
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
December 2009