Barstow, David A. (1983) Bacillus subtilis and bacillus licheniformis as hosts for genetic manipulation. PhD thesis, University of Warwick.

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Abstract

Because of the potential use of the Bacilli for genetic manipulation, experiments were undertaken to investigate the usefulness of Bacillus subtilis and Bacillus licheniformis strain LO2, as hosts.

Attempts at shotgun-cloning directly in B. subtilis met with repeated failure. However, subsequently the Φ3T-thyP3 gene, pC194 chloramphenicol acetyl transferase gene, B. licheniformis 749/C penP gene and the E. coli lacZ gene were expressed in B. subtilis when cloned into the plasmid vector pAB224 or one of its derivatives. Consequently, such plasmids are useful vectors for genetic manipulation in B. subtilis.

The properties of the thyP3-containing hybrids were investigated. Of particular interest was the finding that monomeric plasmid DNA, containing the thyP3 gene, was active in the transformation of competent B. subtilis cells. Transformation resulted in integration of the thyP3-containing region of the plasmid into the host chromosome.

The secretion of fusion proteins by B. subtilis was investigated employing the B. licheniformis 749/C penicillinase protein signal-peptide. This resulted in the secretion of the E. coli β-galactosidase enzyme from the B. subtilis cell.

The thermotolerant Bacillus licheniformis strain LO2 was investigated as a possible host for genetic manipulation. A series of mutant strains were isolated but the induction of competence in two such strains could not be achieved. Additionally, transformation of protoplasts of strain LO2 could not be demonstrated. Thus contrary to previous hopes, at present this strain does not appear to be suitable as a host for genetic manipulation.

Item Type:	Thesis (PhD)
Subjects:	Q Science > QH Natural history > QH426 Genetics
Library of Congress Subject Headings (LCSH):	Genetic engineering, Bacterial genetics, Bacillus (Bacteria) -- Genetics, Plasmids -- Genetics
Official Date:	November 1983
Dates:	Date Event November 1983 Submitted
Institution:	University of Warwick
Theses Department:	Department of Biological Sciences
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Atkinson, A. ; Primrose, S. B.
Sponsors:	Great Britain. Health and Safety Executive
Format of File:	pdf
Extent:	xiv, 255 leaves : illustrations, charts
Language:	eng

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