Document Detail


Model suicide vector for containment of genetically engineered microorganisms.
MedLine Citation:
PMID:  3060017     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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)
Authors:
A K Bej; M H Perlin; R M Atlas
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  54     ISSN:  0099-2240     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  1988 Oct 
Date Detail:
Created Date:  1989-01-25     Completed Date:  1989-01-25     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2472-7     Citation Subset:  IM    
Affiliation:
Department of Biology, University of Louisville, Kentucky 40292.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics*
Blotting, Southern
Carbenicillin / pharmacology
Cloning, Molecular
Colony Count, Microbial
DNA Probes
DNA, Bacterial / genetics
Escherichia coli / drug effects,  genetics*
Gene Expression Regulation
Genes, Bacterial*
Genetic Vectors*
Models, Genetic
Mutation
Nucleic Acid Hybridization
Penicillin Resistance
Peptides / genetics
Plasmids*
Transformation, Bacterial
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA Probes; 0/DNA, Bacterial; 0/Peptides; 4697-36-3/Carbenicillin
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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