Construction of Multicopy Expression Vectors for Regulated Over-production of Proteins in Klebsiella pneumoniae and Other Enteric Bacteria

• Published in: Journal of general microbiology Vol. 134; no. 7; pp. 1779 - 1784
• Main Authors: Kleiner, Diethelm, Paul, Wyatt, Merrick, Mike J
• Format: Journal Article
• Language: English
• Published: London Soc General Microbiol 01-07-1988; New York, NY Cambridge University Press
• Subjects: Bacterial Proteins - biosynthesis; Bacteriology; Biological and medical sciences; Biotechnology; Escherichia coli; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Genes, Bacterial; Genetic engineering; Genetic technics; Genetics; Klebsiella pneumoniae - genetics; Klebsiella pneumoniae - metabolism; Methods. Procedures. Technologies; Microbiology; Plasmids; Promoter Regions, Genetic; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Genetic mapping; Enzymatic synthesis; Gene dosage; Transcription promoter; Activation; Gene expression; Genetic transfer; Plasmid; Proteins; Regulation(control); Production; Bacteria; Molecular cloning; Vector; Klebsielleae; Hybrid gene; Klebsiella pneumoniae; Enterobacteriaceae
• ISSN: 0022-1287; 1350-0872; 1465-2080
• DOI: 10.1099/00221287-134-7-1779

AFRC-IPSR Nitrogen Fixation Laboratory, University of Sussex, Brighton BN1 9RQ, UK
ABSTRACT
Summary: A number of expression vectors have been constructed to allow over-production of selected gene products in Klebsiella pneumoniae and other enteric bacteria. The plasmids use the strong hybrid trp-lac ( tac ) promoter for gene expression, which is regulated by the lacI Q allele of the lac repressor carried on the vector. This provides very tight regulation of gene expression, which is important for over-production of proteins which may be detrimental to cell growth. The vectors carry the standard mpl8 cloning nest in which all the restriction sites are unique to the plasmid (with the exception of Eco RI in pDK7). Derivatives were constructed carrying kanamycin, chloramphenicol or ampillicin resistance as selectable markers, the first two of which are advantageous in K. pneumoniae due to the high inherent β-lactamase activity of this organism.
Present address: Lehrstuhl für Mikrobiologie, Universität Bayreuth, 8580 Bayreuth, FRG.