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 Bej, A K Articles by Atlas, R M Search for Related Content PubMed PubMed Citation Articles by Bej, A K Articles by Atlas, R M Agricola Articles by Bej, A K Articles by Atlas, R M

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1988 October; 54(10): 2472-2477

Model suicide vector for containment of genetically engineered microorganisms.

Department of Biology, University of Louisville, Kentucky 40292.

ABSTRACT

A model suicide vector (pBAP19h), designed for the potential containment of genetically engineered microorganisms, was made by constructing a plasmid with the hok gene, which codes for a lethal polypeptide, under the control of the lac promoter. The vector plasmid also codes for carbenicillin resistance. In the absence of carbenicillin, induction of the hok gene in vitro caused elimination of all detectable cells containing the suicide vector; pBAP19h-free cells of the culture survived and grew exponentially. In the presence of carbenicillin, however, the number of cells containing pBAP19h initially declined after induction of hok but then multiplied exponentially. The surviving cells still had a fully functional hok gene and had apparently developed resistance to the action of the Hok polypeptide. Thus, high selective pressure against the loss of the suicide vector led to a failure of the system. Soil microcosm experiments confirmed the ability of a suicide vector to restrict the growth of a genetically engineered microorganism in the absence of selective pressure against the loss of the plasmid, with 90 to 99% elimination of hok-bearing cells within 24 h of hok induction. However, some pBAP19h-bearing cells survived in the soil microcosms after hok induction. The surviving cells contained an active hok gene but were not capable of normal growth even after elimination of the hok gene; it appears that a mutation that made them Hok resistant also reduced their capacity for membrane functions needed for energy generation and exponential cell growth. Thus, the model suicide vector was shown to be functional in soil as well as in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

• Torres, B., Jaenecke, S., Timmis, K. N., Garcia, J. L., Diaz, E. (2003). A dual lethal system to enhance containment of recombinant micro-organisms. Microbiology 149: 3595-3601 [Abstract] [Full Text]  
• Lindow, S., Panopoulos, N., McFarland, B. (1989). Genetic engineering of bacteria from managed and natural habitats. Science 244: 1300-1307 [Abstract]