Brehm, John Karl, 1959- (1989) Studies towards the development of a host:vector system for the genus Clostridium. PhD thesis, University of Warwick.

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Abstract

Clostridia are widely recognised as organisms of biotechnological importance. This potential, however, cannot be fully exploited until reliable methods have been developed for the transfer of genetic information into and between members of the genus. As with other Gram-positive bacteria, attempts to develop host:vector systems have focused on protoplast transformation procedures. This approach, however, has been hampered by the lack of suitable plasmid vectors. Studies have been initiated to construct such a vector.

As a potential source of a clostridial replicon, the cryptic plasmid of C. butvricum NCIB 7423 (pCBlOl, 6.05 kb) has been examined. The complete nucleotide sequence of pCBlOl was determined and it's minimal replicon characterised together with the elucidation of some of the mechanisms involved in it's replication.

The erythromycin resistance determinant (Emr) of the Grampositive R-factor pAM/3l was chosen as a selectable marker and the entire nucleotide sequence of this gene was determined. The Ferredoxin (Fd) gene of Clostridium pasteurianum was cloned and employed in the construction of an expression cartridge. The effectiveness of this cartridge in promoting the expression of heterologous genes was examined using a promoter-less xvlE gene.

The conjugative Streptococcus faecalis plasmid pAM/Jl was used to mobilise vectors carrying the xvlE gene (under the transcriptional control of the Fd promoter) into Clostridium acetobutvlicum where the gene was expressed. This represents one of the first reports of directed gene expression in ç, acetqbutyllgmn-

Item Type:	Thesis or Dissertation (PhD)
Subjects:	Q Science > QR Microbiology T Technology > TP Chemical technology
Library of Congress Subject Headings (LCSH):	Clostridium, Clostridium -- Biotechnology, Microbial biotechnology
Official Date:	April 1989
Dates:	Date Event April 1989 UNSPECIFIED
Institution:	University of Warwick
Theses Department:	Department of Biological Sciences
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Minton, Nigel P. ; Salmond, George
Format of File:	pdf
Extent:	xvi, 289 leaves : illustrations
Language:	eng

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