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Applied and Environmental Microbiology, November 1998, p. 4618-4622, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Design of a Phage-Insensitive Lactococcal Dairy Starter via Sequential Transfer of Naturally Occurring Conjugative Plasmids

David O'Sullivan,1,2 Aidan Coffey,1 Gerald F. Fitzgerald,2 Colin Hill,2 and R. Paul Ross1,*

Dairy Products Research Centre, Teagasc, Moorepark, Fermoy, County Cork,1 and Department of Microbiology, University College, Cork,2 Ireland

Received 3 April 1998/Accepted 18 August 1998

The plasmid-free Lactococcus lactis subsp. cremoris MG1614 is highly phage sensitive and lacks lactose fermenting ability (Lac) and primary casein degrading ability (Prt). Food grade gene transfer systems were used to sequentially superimpose different phage defense systems on this background, resulting in a gradual increase in resistance to bacteriophage in the derivatives. pLP712, encoding Lac and Prt, was then transferred to one of these hosts, into which plasmids encoding adsorption inhibition, restriction modification, and abortive infection had already been introduced. This resulted in a phage-resistant strain which was successfully used as a single-strain starter for cheddar cheese manufacture under industrial conditions.

* Corresponding author. Mailing address: Dairy Products Research Centre, Teagasc, Moorepark, Fermoy, Co. Cork, Ireland. Phone: 353-25-42222. Fax: 353-25-42340. E-mail: pross{at}moorepark.teagasc.ie.

Applied and Environmental Microbiology, November 1998, p. 4618-4622, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • O'Sullivan, L., Ryan, M. P., Ross, R. P., Hill, C. (2003). Generation of Food-Grade Lactococcal Starters Which Produce the Lantibiotics Lacticin 3147 and Lacticin 481. Appl. Environ. Microbiol. 69: 3681-3685 [Abstract] [Full Text]  
  • Ryan, M. P., Ross, R. P., Hill, C. (2001). Strategy for Manipulation of Cheese Flora Using Combinations of Lacticin 3147-Producing and -Resistant Cultures. Appl. Environ. Microbiol. 67: 2699-2704 [Abstract] [Full Text]  
  • Hickey, R. M., Twomey, D. P., Ross, R. P., Hill, C. (2001). Exploitation of Plasmid pMRC01 To Direct Transfer of Mobilizable Plasmids into Commercial Lactococcal Starter Strains. Appl. Environ. Microbiol. 67: 2853-2858 [Abstract] [Full Text]  
  • McGrath, S., Fitzgerald, G. F., van Sinderen, D. (2001). Improvement and Optimization of Two Engineered Phage Resistance Mechanisms in Lactococcus lactis. Appl. Environ. Microbiol. 67: 608-616 [Abstract] [Full Text]  
  • O' Sullivan, D., Ross, R. P., Twomey, D. P., Fitzgerald, G. F., Hill, C., Coffey, A. (2001). Naturally Occurring Lactococcal Plasmid pAH90 Links Bacteriophage Resistance and Mobility Functions to a Food-Grade Selectable Marker. Appl. Environ. Microbiol. 67: 929-937 [Abstract] [Full Text]  
  • O'Sullivan, D., Ross, R. P., Fitzgerald, G. F., Coffey, A. (2000). Investigation of the Relationship between Lysogeny and Lysis of Lactococcus lactis in Cheese Using Prophage-Targeted PCR. Appl. Environ. Microbiol. 66: 2192-2198 [Abstract] [Full Text]  
  • Walker, S. A., Klaenhammer, T. R. (2000). An Explosive Antisense RNA Strategy for Inhibition of a Lactococcal Bacteriophage. Appl. Environ. Microbiol. 66: 310-319 [Abstract] [Full Text]  


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