Publication Date

1981

Document Type

Dissertation/Thesis

First Advisor

Vary, Patricia S.

Degree Name

M.S. (Master of Science)

Legacy Department

Department of Biological Sciences

LCSH

Bacterial genetics; Leucine; Bacillus megaterium; Amino acids--Synthesis

Abstract

This thesis represents work done to determine the optimal conditions for transduction of Bacillus megaterium QM B1551 by bacteriophage MP13 and to use MP13 to initiate genetic studies of leucine biosynthesis in this bacterium. To determine the conditions which would increase the proportion of transducing particles, media, pH, temperature, aeration, multiplicity of infection and growth stage of the host were varied during propagation of the lysate. Conditions of the transduction procedure which were varied included: growth media of the recipient, phage inactivation time, selective media, pH of the selective media, incubation temperature and the use of antiserum. The results indicated a higher proportion of transducing particles were produced when the lysates were propagated in SNB broth, pH 8.0 at 30°C or 37°C with reduced aeration and a low multiplicity of infection. Recovery of transductants was increased when the recipient was grown in M broth at 30°C to late logarithmic stage, and the recipient was plated on an acidic selection medium with phage inactivated by 20 to 40 seconds of UV exposure. Increased recovery of transductants was observed when incubation was at 30°C but antiserum did not enhance recovery. After establishment of these optimal conditions for transduction, MP13 was used to analyse the leucine genes of 13. megaterium. Ten leucine mutations were found to be cotransducible and linked to ilv and phe markers. Biochemical and genetic analysis indicated the order of the genes to be ilv-leuA-leuC-leuB-phe.

Comments

Includes bibliographical references.

Extent

80 pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

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