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Applied and Environmental Microbiology, August 2005, p. 4897-4901, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4897-4901.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Insertional Inactivation of Determinants for Mg2+ and Co2+ Transport as a Tool for Screening Recombinant Lactococcus Species Clones

Susan Mills,1,2 Aidan Coffey,3 Colin Hill,2,4 Gerald F. Fitzgerald,2,4 Olivia McAuliffe,1* and R. Paul Ross1,4

Teagasc, Dairy Products Research Centre, Moorepark, Fermoy, County Cork, Ireland,1 Department of Microbiology, University College Cork,2 Alimentary Pharmabiotic Centre,4 Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland3

Received 21 February 2005/ Accepted 28 February 2005

Insertional inactivation of the plasmid-encoded determinants for Mg2+ and Co2+ transport, orf18/corA, provides a tool for screening recombinant clones in Lactococcus, based on the observation that overexpression of orf18/corA results in cell growth inhibition on certain concentrations of CoCl2. The lacticin 3147 immunity gene, ltnI, was used to insertionally inactivate orf18/corA. The resulting clones were capable of growth on concentrations of CoCl2 that were inhibitory to the parent strain. Since only 3 of 17 lactococcal starters naturally harbor corA, the system has potential as a screen for selecting recombinant lactococcal clones.

* Corresponding author. Mailing address: Teagasc, Dairy Products Research Centre, Moorepark, Fermoy, County Cork, Ireland. Phone: 353-25-42609. Fax: 353-25-42340. E-mail: omcauliffe{at}moorepark.teagasc.ie.

Applied and Environmental Microbiology, August 2005, p. 4897-4901, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4897-4901.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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