Abstract
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 microorganism 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.
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Munthali, M., Timmis, K. & Díaz, E. Restricting the Dispersal of Recombinant DNA: Design of a Contained Biological Catalyst. Nat Biotechnol 14, 189–191 (1996). https://doi.org/10.1038/nbt0296-189
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DOI: https://doi.org/10.1038/nbt0296-189