BIOINFORMATIC ANALYSIS OF A MAMMALIAN BIP GENE FOR INSERTION INTO GREEN ALGAE AND COMPARISON OF ITS POSSIBLE EFFECTS ON THE SYNTHESIS OF A MAMMALIAN ANTIBODY

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.

Name thesis:degree_name

Doctor of Philosophy

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology & Immunology

Year dc:date.available

2004

Author dc:creator

• Ghazanfar, Katrina

Contributors dc:contributor

• Fang-Sheng Wu
• Guy Cabral
• Kathleen McCoy
• Jennifer Stewart
• L. Shozo Ozaki

• BIP
• Bioinformatics
• HuCC49
• Chlamydomonas reinhardtii
• GRP 78
• pSP124S
• electroporation
• plasmolysis
• Medicine and Health Sciences

Statement dc:rights

• © The Author

Identifier

https://scholarscompass.vcu.edu/etd/36

OAI identifier oai:identifier

oai:scholarscompass.vcu.edu:etd-1035