Restricting the dispersal of recombinant DNA: design of a contained biological catalyst.
To restrict horizontal gene spread and, thus, create organisms whose behavior in the field is more predictable, we have combined a mini-Tn5 cloning system that allows stable insertion of foreign genes into the chromosomes of a variety of Gram-negative bacteria with a gene-containment circuit based on the universal lethal-function colicin E3. Use of the system is exemplified by the construction of a micro-organism designed to degrade polychlorinated biphenyls, important environmental pollutants. The relevant genotype of the microorganism is subject to a stringent gene-containment mechanism that provides neither an advantage nor a disadvantage to the host cell for survival, but decreases frequencies of productive chromosomal transfer frequencies by at least four orders of magnitude.
Munthal
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MT
Department of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig, Germany.
Timmis
K N
KN
Díaz
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D002384:Catalysis; D003087:Colicins / Q000235:genetics; D003264:Containment of Biohazards; D004251:DNA Transposable Elements; D004274:DNA, Recombinant / Q000235:genetics*; D022761:Gene Transfer, Horizontal; D005818:Genetic Engineering; D008040:Genetic Linkage; D006090:Gram-Negative Bacteria / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016958:Pseudomonas putida / Q000235:genetics
D003087:Colicins; D004251:DNA Transposable Elements; D004274:DNA, Recombinant