Construction of a Candida albicans genomic library and isolation of a gene which complements B-isopropylmalate dehydrogenase mutations in Esherichia coli and Saccharomyces cerevisiae / by Michael Aaron Turnage.
| Author/creator | Turnage, Michael Aaron author. |
| Other author | Baptist, Edward W., degree supervisor. |
| Other author | Kennedy, Kathleen E., 1946- degree supervisor. |
| Other author | East Carolina University. Department of Biology. |
| Format | Theses and dissertations |
| Production | 1989. |
| Description | [iii], 66 leaves : illustrations ; 28 cm |
| Supplemental Content | Access via ScholarShip |
| Subjects |
| Summary | The initial goal of this research project was to isolate and characterize the Candida albicans gene for dihydrofolate reductase (DHFR) by constructing a genomic library from DNA isolated from this yeast in Escherichia coli and screening the library for clones which confer to E. coli resistance to trimethoprim, a drug which inhibits the bacterial DHFR but is not inhibitory to the enzyme produced by many eukaryotes. The DHFR gene was not isolated by the methods employed, however a plasmid containing a fragment of C. albicans DNA was isolated which complements an E. coli mutation in the gene for [beta]-isopropyl malate dehydrogenase (leuB). This plasmid was used to transform the yeast Saccharamyces cerevisiae and found to complement the corresponding mutation (leu2) in this organism. Approximately 10,000 individual plasmids were isolated, tested for the presence of the DHFR gene, and stored for future use. Analysis of DNA from 100 randomly selected clones indicated that the average size fragment of yeast DNA contained was at least 5000 base pairs, giving a probability of 95% that any sequence in the yeast genome is present at least once in the library. |
| General note | Submitted to the faculty of the Department of Biology. |
| General note | Advisor: Edward W. Baptist |
| General note | Advisor: Kathleen E. Kennedy |
| Dissertation note | M.S. East Carolina University 1989 |
| Bibliography note | Includes bibliographical references (leaves 62-66). |
| Genre/form | Academic theses. |
| Genre/form | Academic theses. |
| Genre/form | Thèses et écrits académiques. |
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