Thymidylate synthase gene from Lactococcus lactis as a genetic marker: an alternative to antibiotic resistance genes.

Applied and Environmental Microbiology
P RossS Condon

Abstract

The potential of the thymidylate synthase thyA gene cloned from Lactococcus lactis subsp. lactis as a possible alternative selectable marker gene to antibiotic resistance markers has been examined. The thyA mutation is a recessive lethal one; thyA mutants cannot survive in environments containing low amounts of thymidine or thymine (such as Luria-Bertani medium) unless complemented by the thyA gene. The cloned thyA gene was strongly expressed in L. lactis subsp. lactis, Escherichia coli, Rhizobium meliloti, and a fluorescent Pseudomonas strain. In addition, when fused to a promoterless enteric lac operon, the thyA gene drove expression of the lac genes in a number of gram-negative bacteria. In transformation experiments with thyA mutants of E. coli and conjugation experiments with thyA mutants of R. meliloti, the lactococcal thyA gene permitted selection of transformants and transconjugants with the same efficiency as did genes for resistance to ampicillin, chloramphenicol, or tetracycline. Starting from the broad-host-range plasmid pGD500, a plasmid, designated pPR602, was constructed which is completely free of antibiotic resistance genes and has the lactococcal thyA gene fused to a promoterless lac operon. This plasmid will ...Continue Reading

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Citations

Jul 6, 2000·Applied and Environmental Microbiology·S W Lee, D A Cooksey
Jan 27, 2019·Applied and Environmental Microbiology·Laura M AlexanderJan-Peter van Pijkeren
Jul 1, 1996·Molecular and Biochemical Parasitology·R ArrebolaD González-Pacanowska
Nov 1, 1992·Journal of Bacteriology·P Le BourgeoisP Ritzenthaler
Jul 1, 1990·Applied and Environmental Microbiology·P RossS Condon
Jun 25, 2015·Microbiology Spectrum·Esteban Martínez-GarcíaVíctor De Lorenzo
Mar 1, 1996·Applied and Environmental Microbiology·C PlatteeuwW M de Vos

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