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 Ahrenholtz, I Articles by Wackernagel, W Search for Related Content PubMed PubMed Citation Articles by Ahrenholtz, I Articles by Wackernagel, W Agricola Articles by Ahrenholtz, I Articles by Wackernagel, W

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1994 October; 60(10): 3746-3751

A conditional suicide system in Escherichia coli based on the intracellular degradation of DNA.

Universität Oldenburg, Germany.

ABSTRACT

The potential risks associated with the intentional or unintentional release of genetically engineered microorganisms led to the construction of biological containment systems by which bacteria are killed in a controlled suicide process. In previously published suicide systems, cell killing was caused by proteins destroying the cell membrane or cell wall. Here a conditional cell killing system based on the intracellular degradation of cellular DNA is presented. The nuclease gene used was that of the extracellular nuclease of Serratia marcescens. The nuclease gene was deleted for the leader-coding sequence, and the truncated gene was put under the control of the lambda pL promoter. Following thermoinduction of the nuclease gene cassette in Escherichia coli, cell survival dropped to 2 x 10(-5), and more than 80% of the radioactively labeled DNA was converted to acid-soluble material within 2.5 h in the absence of cell lysis. The majority (84%) of clones which survived thermoinduced killing turned out to be as sensitive to a second thermoinduction as the original strain. The other clones showed somewhat slower killing kinetics or slightly higher final levels of survivors. The suicide system described combines the regulated killing of cells with the destruction of intracellular DNA otherwise potentially available for horizontal gene transfer processes.

This article has been cited by other articles:

• Haidinger, W., Mayr, U. B., Szostak, M. P., Resch, S., Lubitz, W. (2003). Escherichia coli Ghost Production by Expression of Lysis Gene E and Staphylococcal Nuclease. Appl. Environ. Microbiol. 69: 6106-6113 [Abstract] [Full Text]  
• Sakamoto, J. J., Sasaki, M., Tsuchido, T. (2001). Purification and Characterization of a Bacillus subtilis 168 Nuclease, YokF, Involved in Chromosomal DNA Degradation and Cell Death Caused by Thermal Shock Treatments. J. Biol. Chem. 276: 47046-47051 [Abstract] [Full Text]  
• Le Loir, Y., Nouaille, S., Commissaire, J., Bretigny, L., Gruss, A., Langella, P. (2001). Signal Peptide and Propeptide Optimization for Heterologous Protein Secretion in Lactococcus lactis. Appl. Environ. Microbiol. 67: 4119-4127 [Abstract] [Full Text]  
• Poquet, I., Ehrlich, S. D., Gruss, A. (1998). An Export-Specific Reporter Designed for Gram-Positive Bacteria: Application to Lactococcus lactis. J. Bacteriol. 180: 1904-1912 [Abstract] [Full Text]  
• Szafranski, P., Mello, C. M., Sano, T., Smith, C. L., Kaplan, D. L., Cantor, C. R. (1997). A new approach for containment of microorganisms: Dual control of streptavidin expression by antisense RNA and the T7 transcription system. Proc. Natl. Acad. Sci. USA 94: 1059-1063 [Abstract] [Full Text]