Comparative intragenic mapping of aspartic-acid auxotrophs of Staphylococcus Aureus using transduction and transformation / by James Van Taylor, III.

Author/creator Van Taylor, James author.
Other author Allen, Wendall E., degree supervisor.
Other author East Carolina University. Department of Biology.
Format Theses and dissertations
Production1976.
Description68 leaves : illustrations ; 29 cm
Supplemental ContentAccess via ScholarShip
Subjects

Summary Twenty-nine DNase⁻ auxotrophic strains obtained after mutagenic treatment of the Staphylococcus aureus parent strain 7-8 with N-methyl-Ń- nitro-K-nitrosoguanidine were selected for use in fine-structure genetic analysis. The auxotrophs were divided into two groups, those that showed evidence of reversion, probably representing point mutations, and those that showed no evidence of reversion, probably representing deletion or multisite mutations. Auxotrophs having point mutations were proposed to have chromosomal defects within the same genetic locus as determined by the lack of syntrophism. The auxotrophs having point mutations were tested for lytic sensitivity to the serological group B staphylococcal typing phages 52, 53, 29, 79, 80, 81 and phage Ø11. Auxotrophs and their respective lytic phages were designated for possible use in transductlonal studies. A transductlonal screening test was used to identify transductionally competent phageauxotroph groups for use in fine-structure genetic analysis. Each of the staphylophages showed competency in transducing some portion of the auxotrophs. The aspartic-acid-deflcient auxotrophs, Asp-⁻4, Asp-⁻7, and Asp-⁻11 were selected for transductlonal analysis using typing phage 80 and phage Ø11 and for transformational analysis, Lysogeny was shown not to be a requisite for the aspartic-acid transductions, Transductional frequencies of the phage Ø11-lysogenized mutants [Asp-⁻4 (Ø11) , Asp-⁻7 (Ø11), and Asp-~11 (Ø11)] using typing phage 80, were lower than the transductional frequencies using the non-lysogenized auxotrophs and the same phage. Phage Ø11 was not as effective a transducing phage for the non-lysogenized auxotrophs as typing phage 80, Intragenic maps of three aspartic-acid mutation sites on the chromosome of S. aureus were prepared from data obtained by three methods: reciprocaltransduction analysis using typing phage 80 and phage Ø11, recipientcapacity determination using typing phage 80, and transformational analysis of the phage Ø11-lysogenized mutants. There was agreement in the order and relative position of the three aspartic-acid mutation sites determined from results of reciprocal-transduction experiments using typing phage 80 and recipient-capacity experiments. Results of preliminary experiments for both transductional analysis with phage Ø11 and transformational analysis of these mutants were inconclusive.
Local noteJoyner-"Presented to the faculty of the Department of Biology ... in partial fulfillment of the requirements for the degree Master of Science in Biology."
General notePresented to the faculty of the Department of Biology.
General noteAdvisor: Wendall E. Allen
Dissertation noteM.S. East Carolina University 1976
Bibliography noteIncludes bibliographical references (leaves 61-68).
Genre/formdissertations.
Genre/formAcademic theses.
Genre/formAcademic theses.
Genre/formThèses et écrits académiques.

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