This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by O'Connor, C D Articles by Timmis, K N Search for Related Content PubMed PubMed Citation Articles by O'Connor, C D Articles by Timmis, K N

 Previous Article  |  Next Article 

J Bacteriol. 1987 October; 169(10): 4457-4462

Highly repressible expression system for cloning genes that specify potentially toxic proteins.

Department of Medical Biochemistry, University of Geneva, Switzerland.

ABSTRACT

A highly repressible expression vector system that allows the cloning of potentially deleterious genes has been constructed. Undesired expression of a cloned gene was prevented (i) at the level of initiation of transcription, by the presence of the strong but highly repressible leftward promoter of bacteriophage lambda, lambda pL, and (ii) at the level of transcript elongation or translation, through synthesis of antisense RNA complementary to the mRNA of the cloned gene. The system was tested by measuring the inhibition of expression of traT, the gene for the TraT major outer membrane lipoprotein. Direct detection and functional assays indicated that an essentially complete inhibition of traT expression was obtained. As a further test of the system, the gene encoding the EcoRI restriction endonuclease was cloned in the absence of the gene of the corresponding protective EcoRI modification methylase. Transformants harboring this construct were only viable when both repression controls were operational.

This article has been cited by other articles:

• Takac, M., Witte, A., Blasi, U. (2005). Functional analysis of the lysis genes of Staphylococcus aureus phage P68 in Escherichia coli. Microbiology 151: 2331-2342 [Abstract] [Full Text]  
• Kong, H., Lin, L.-F., Porter, N., Stickel, S., Byrd, D., Posfai, J., Roberts, R. J. (2000). Functional analysis of putative restriction-modification system genes in the Helicobacter pylori J99 genome. Nucleic Acids Res 28: 3216-3223 [Abstract] [Full Text]