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
Production1989.
Description[iii], 66 leaves : illustrations ; 28 cm
Supplemental ContentAccess 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 noteSubmitted to the faculty of the Department of Biology.
General noteAdvisor: Edward W. Baptist
General noteAdvisor: Kathleen E. Kennedy
Dissertation noteM.S. East Carolina University 1989
Bibliography noteIncludes bibliographical references (leaves 62-66).
Genre/formAcademic theses.
Genre/formAcademic theses.
Genre/formThèses et écrits académiques.

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