This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Breüner, A.
Right arrow Articles by Hammer, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Breüner, A.
Right arrow Articles by Hammer, K.

 Previous Article  |  Next Article 

Journal of Bacteriology, December 1999, p. 7291-7297, Vol. 181, No. 23
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Novel Organization of Genes Involved in Prophage Excision Identified in the Temperate Lactococcal Bacteriophage TP901-1

Anne Breüner, Lone Brøndsted, and Karin Hammer*

Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark

Received 1 June 1999/Accepted 17 September 1999

In this work, the phage-encoded proteins involved in site-specific excision of the prophage genome of the temperate lactococcal bacteriophage TP901-1 were identified. The phage integrase is required for the process, and a low but significant frequency of excision is observed when the integrase is the only phage protein present. However, 100% excision is observed when the phage protein Orf7 is provided as well as the integrase. Thus, Orf7 is the TP901-1 excisionase, and it is the first excisionase identified that is used during excisive recombination catalyzed by an integrase belonging to the family of extended resolvases. Orf7 is a basic protein of 64 amino acids, and the corresponding gene (orf7) is the third gene in the early lytic operon. This location of an excisionase gene of a temperate bacteriophage has never been described before. The experiments are based on in vivo excision of specifically designed excision vectors carrying the TP901-1 attP site which are integrated into attB on the chromosome of Lactococcus lactis. Excision of the vectors was investigated in the presence of different TP901-1 genes. In order to detect very low frequencies of excision, a method for positive selection of loss of genetic material based upon the upp gene (encoding uracil phosphoribosyltransferase) was designed, since upp mutants are resistant to fluorouracil. By using this system, frequencies of excision on the order of 10-5 per cell could easily be measured. The described selection principle may be of general use for many organisms and also for types of deletion events other than excision.

* Corresponding author. Mailing address: Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45 45 25 24 96. Fax: 45 45 88 26 60. E-mail: imkh{at}pop.dtu.dk.

Journal of Bacteriology, December 1999, p. 7291-7297, Vol. 181, No. 23
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Haaber, J., Rousseau, G. M., Hammer, K., Moineau, S. (2009). Identification and Characterization of the Phage Gene sav, Involved in Sensitivity to the Lactococcal Abortive Infection Mechanism AbiV. Appl. Environ. Microbiol. 75: 2484-2494 [Abstract] [Full Text]  
  • Denou, E., Pridmore, R. D., Ventura, M., Pittet, A.-C., Zwahlen, M.-C., Berger, B., Barretto, C., Panoff, J.-M., Brussow, H. (2008). The Role of Prophage for Genome Diversification within a Clonal Lineage of Lactobacillus johnsonii: Characterization of the Defective Prophage LJ771. J. Bacteriol. 190: 5806-5813 [Abstract] [Full Text]  
  • Solem, C., Defoor, E., Jensen, P. R., Martinussen, J. (2008). Plasmid pCS1966, a New Selection/Counterselection Tool for Lactic Acid Bacterium Strain Construction Based on the oroP Gene, Encoding an Orotate Transporter from Lactococcus lactis. Appl. Environ. Microbiol. 74: 4772-4775 [Abstract] [Full Text]  
  • Ghosh, P., Bibb, L. A., Hatfull, G. F. (2008). Two-step site selection for serine-integrase-mediated excision: DNA-directed integrase conformation and central dinucleotide proofreading. Proc. Natl. Acad. Sci. USA 105: 3238-3243 [Abstract] [Full Text]  
  • Defoor, E., Kryger, M.-B., Martinussen, J. (2007). The orotate transporter encoded by oroP from Lactococcus lactis is required for orotate utilization and has utility as a food-grade selectable marker. Microbiology 153: 3645-3659 [Abstract] [Full Text]  
  • Vegge, C. S., Brondsted, L., Neve, H., Mc Grath, S., van Sinderen, D., Vogensen, F. K. (2005). Structural Characterization and Assembly of the Distal Tail Structure of the Temperate Lactococcal Bacteriophage TP901-1. J. Bacteriol. 187: 4187-4197 [Abstract] [Full Text]  
  • Lucet, I. S., Tynan, F. E., Adams, V., Rossjohn, J., Lyras, D., Rood, J. I. (2005). Identification of the Structural and Functional Domains of the Large Serine Recombinase TnpX from Clostridium perfringens. J. Biol. Chem. 280: 2503-2511 [Abstract] [Full Text]  
  • Smith, M. C. A., Till, R., Brady, K., Soultanas, P., Thorpe, H., Smith, M. C. M. (2004). Synapsis and DNA cleavage in {phi}C31 integrase-mediated site-specific recombination. Nucleic Acids Res 32: 2607-2617 [Abstract] [Full Text]  
  • Canchaya, C., Proux, C., Fournous, G., Bruttin, A., Brussow, H. (2003). Prophage Genomics. Microbiol. Mol. Biol. Rev. 67: 238-276 [Abstract] [Full Text]  
  • Proux, C., van Sinderen, D., Suarez, J., Garcia, P., Ladero, V., Fitzgerald, G. F., Desiere, F., Brussow, H. (2002). The Dilemma of Phage Taxonomy Illustrated by Comparative Genomics of Sfi21-Like Siphoviridae in Lactic Acid Bacteria. J. Bacteriol. 184: 6026-6036 [Abstract] [Full Text]  
  • Combes, P., Till, R., Bee, S., Smith, M. C. M. (2002). The Streptomyces Genome Contains Multiple Pseudo-attB Sites for the {phi}C31-Encoded Site-Specific Recombination System. J. Bacteriol. 184: 5746-5752 [Abstract] [Full Text]  
  • Stoll, S. M., Ginsburg, D. S., Calos, M. P. (2002). Phage TP901-1 Site-Specific Integrase Functions in Human Cells. J. Bacteriol. 184: 3657-3663 [Abstract] [Full Text]  
  • Crutz-Le Coq, A.-M., Cesselin, B., Commissaire, J., Anba, J. (2002). Sequence analysis of the lactococcal bacteriophage bIL170: insights into structural proteins and HNH endonucleases in dairy phages. Microbiology 148: 985-1001 [Abstract] [Full Text]  
  • Breuner, A., Hammer, K. (2001). Resolvase-like recombination performed by the TP901-1 integrase. Microbiology 147: 2051-2063 [Abstract] [Full Text]  
  • Lewis, J. A., Hatfull, G. F. (2001). Control of directionality in integrase-mediated recombination: examination of recombination directionality factors (RDFs) including Xis and Cox proteins. Nucleic Acids Res 29: 2205-2216 [Abstract] [Full Text]  
  • Madsen, P. L., Johansen, A. H., Hammer, K., Brøndsted, L. (1999). The Genetic Switch Regulating Activity of Early Promoters of the Temperate Lactococcal Bacteriophage TP901-1. J. Bacteriol. 181: 7430-7438 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»