This Article
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 Rosa, P.
Right arrow Articles by Tilly, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rosa, P.
Right arrow Articles by Tilly, K.

 Previous Article  |  Next Article 

J. Bacteriol., Oct 1996, 5946-5953, Vol 178, No. 20
Copyright © 1996, American Society for Microbiology

Directed insertion of a selectable marker into a circular plasmid of Borrelia burgdorferi

P Rosa, DS Samuels, D Hogan, B Stevenson, S Casjens and K Tilly
Laboratory of Microbial Structure and Function, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840, USA. patricia_rosa@nih.gov

Studies of the biology of Borrelia burgdorferi and the pathogenesis of Lyme disease are severely limited by the current lack of genetic tools. As an initial step toward facile genetic manipulation of this pathogenic spirochete, we have investigated gene inactivation by allelic exchange using a mutated borrelial gyrB gene that confers resistance to the antibiotic coumermycin A1 as a selectable marker. We have transformed B. burgdorferi by electroporation with a linear fragment of DNA in which this selectable marker was flanked by sequences from a native borrelial 26-kb circular plasmid. We have identified coumermycin A1-resistant transformants in which gyrB had interrupted the targeted site on the 26-kb plasmid via homologous recombination with the flanking sequences. Antibiotic resistance conferred by the mutated gyrB gene on the plasmid is dominant, and transformed spirochetes carrying this plasmid do not contain any unaltered copies of the plasmid. Coumermycin A1 resistance can be transferred to naive B. burgdorferi by transformation with borrelial plasmid DNA from the initial transformants. This work represents the first example of a directed mutation in B. burgdorferi whereby a large segment of heterologous DNA (gyrB) has been inserted via homologous recombination with flanking sequences, thus demonstrating the feasibility of specific gene inactivation by allelic exchange.


This article has been cited by other articles:

  • Sarkar, A., Tilly, K., Stewart, P., Bestor, A., Battisti, J. M., Rosa, P. A. (2009). Borrelia burgdorferi Resistance to a Major Skin Antimicrobial Peptide Is Independent of Outer Surface Lipoprotein Content. Antimicrob. Agents Chemother. 53: 4490-4494 [Abstract] [Full Text]  
  • Battisti, J. M., Bono, J. L., Rosa, P. A., Schrumpf, M. E., Schwan, T. G., Policastro, P. F. (2008). Outer Surface Protein A Protects Lyme Disease Spirochetes from Acquired Host Immunity in the Tick Vector. Infect. Immun. 76: 5228-5237 [Abstract] [Full Text]  
  • Stewart, P. E., Bestor, A., Cullen, J. N., Rosa, P. A. (2008). A Tightly Regulated Surface Protein of Borrelia burgdorferi Is Not Essential to the Mouse-Tick Infectious Cycle. Infect. Immun. 76: 1970-1978 [Abstract] [Full Text]  
  • Stewart, P. E., Wang, X., Bueschel, D. M., Clifton, D. R., Grimm, D., Tilly, K., Carroll, J. A., Weis, J. J., Rosa, P. A. (2006). Delineating the Requirement for the Borrelia burgdorferi Virulence Factor OspC in the Mammalian Host.. Infect. Immun. 74: 3547-3553 [Abstract] [Full Text]  
  • Pinne, M., Denker, K., Nilsson, E., Benz, R., Bergstrom, S. (2006). The BBA01 Protein, a Member of Paralog Family 48 from Borrelia burgdorferi, Is Potentially Interchangeable with the Channel-Forming Protein P13.. J. Bacteriol. 188: 4207-4217 [Abstract] [Full Text]  
  • Strother, K. O., de Silva, A. (2005). Role of Borrelia burgdorferi Linear Plasmid 25 in Infection of Ixodes scapularis Ticks. J. Bacteriol. 187: 5776-5781 [Abstract] [Full Text]  
  • Stewart, P. E., Hoff, J., Fischer, E., Krum, J. G., Rosa, P. A. (2004). Genome-Wide Transposon Mutagenesis of Borrelia burgdorferi for Identification of Phenotypic Mutants. Appl. Environ. Microbiol. 70: 5973-5979 [Abstract] [Full Text]  
  • Qiu, W.-G., Schutzer, S. E., Bruno, J. F., Attie, O., Xu, Y., Dunn, J. J., Fraser, C. M., Casjens, S. R., Luft, B. J. (2004). Genetic exchange and plasmid transfers in Borrelia burgdorferi sensu stricto revealed by three-way genome comparisons and multilocus sequence typing. Proc. Natl. Acad. Sci. USA 101: 14150-14155 [Abstract] [Full Text]  
  • Byram, R., Stewart, P. E., Rosa, P. (2004). The Essential Nature of the Ubiquitous 26-Kilobase Circular Replicon of Borrelia burgdorferi. J. Bacteriol. 186: 3561-3569 [Abstract] [Full Text]  
  • Ostberg, Y., Carroll, J. A., Pinne, M., Krum, J. G., Rosa, P., Bergstrom, S. (2004). Pleiotropic Effects of Inactivating a Carboxyl-Terminal Protease, CtpA, in Borrelia burgdorferi. J. Bacteriol. 186: 2074-2084 [Abstract] [Full Text]  
  • Frank, K. L., Bundle, S. F., Kresge, M. E., Eggers, C. H., Samuels, D. S. (2003). aadA Confers Streptomycin Resistance in Borrelia burgdorferi. J. Bacteriol. 185: 6723-6727 [Abstract] [Full Text]  
  • Stewart, P. E., Chaconas, G., Rosa, P. (2003). Conservation of Plasmid Maintenance Functions between Linear and Circular Plasmids in Borrelia burgdorferi. J. Bacteriol. 185: 3202-3209 [Abstract] [Full Text]  
  • Miller, J. C., Stevenson, B. (2003). Immunological and genetic characterization of Borrelia burgdorferi BapA and EppA proteins. Microbiology 149: 1113-1125 [Abstract] [Full Text]  
  • Ostberg, Y., Pinne, M., Benz, R., Rosa, P., Bergstrom, S. (2002). Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi. J. Bacteriol. 184: 6811-6819 [Abstract] [Full Text]  
  • Elias, A. F., Stewart, P. E., Grimm, D., Caimano, M. J., Eggers, C. H., Tilly, K., Bono, J. L., Akins, D. R., Radolf, J. D., Schwan, T. G., Rosa, P. (2002). Clonal Polymorphism of Borrelia burgdorferi Strain B31 MI: Implications for Mutagenesis in an Infectious Strain Background. Infect. Immun. 70: 2139-2150 [Abstract] [Full Text]  
  • Tilly, K., Elias, A. F., Errett, J., Fischer, E., Iyer, R., Schwartz, I., Bono, J. L., Rosa, P. (2001). Genetics and Regulation of Chitobiose Utilization in Borrelia burgdorferi. J. Bacteriol. 183: 5544-5553 [Abstract] [Full Text]  
  • Stanton, T. B., Matson, E. G., Humphrey, S. B. (2001). Brachyspira (Serpulina) hyodysenteriae gyrB Mutants and Interstrain Transfer of Coumermycin A1 Resistance. Appl. Environ. Microbiol. 67: 2037-2043 [Abstract] [Full Text]  
  • Thomas, V., Anguita, J., Samanta, S., Rosa, P. A., Stewart, P., Barthold, S. W., Fikrig, E. (2001). Dissociation of Infectivity and Pathogenicity in Borrelia burgdorferi. Infect. Immun. 69: 3507-3509 [Abstract] [Full Text]  
  • Miller, J. C., Bono, J. L., Babb, K., El-Hage, N., Casjens, S., Stevenson, B. (2000). A Second Allele of eppA in Borrelia burgdorferi Strain B31 Is Located on the Previously Undetected Circular Plasmid cp9-2. J. Bacteriol. 182: 6254-6258 [Abstract] [Full Text]  
  • Motaleb, M. A., Corum, L., Bono, J. L., Elias, A. F., Rosa, P., Samuels, D. S., Charon, N. W. (2000). Borrelia burgdorferi periplasmic flagella have both skeletal and motility functions. Proc. Natl. Acad. Sci. USA 10.1073/pnas.200221797v1 [Abstract] [Full Text]  
  • Elias, A. F., Bono, J. L., Carroll, J. A., Stewart, P., Tilly, K., Rosa, P. (2000). Altered Stationary-Phase Response in a Borrelia burgdorferi rpoS Mutant. J. Bacteriol. 182: 2909-2918 [Abstract] [Full Text]  
  • Bono, J. L., Elias, A. F., Kupko, J. J. III, Stevenson, B., Tilly, K., Rosa, P. (2000). Efficient Targeted Mutagenesis in Borrelia burgdorferi. J. Bacteriol. 182: 2445-2452 [Abstract] [Full Text]  
  • Knight, S. W., Kimmel, B. J., Eggers, C. H., Samuels, D. S. (2000). Disruption of the Borrelia burgdorferi gac Gene, Encoding the Naturally Synthesized GyrA C-Terminal Domain. J. Bacteriol. 182: 2048-2051 [Abstract] [Full Text]  
  • Magrini, V., Creighton, C., White, D., Hartzell, P. L., Youderian, P. (1998). The aadA Gene of Plasmid R100 Confers Resistance to Spectinomycin and Streptomycin in Myxococcus xanthus. J. Bacteriol. 180: 6757-6760 [Abstract] [Full Text]  
  • Tilly, K., Lubke, L., Rosa, P. (1998). Characterization of Circular Plasmid Dimers in Borrelia burgdorferi. J. Bacteriol. 180: 5676-5681 [Abstract] [Full Text]  
  • Stevenson, B., Bono, J. L., Elias, A., Tilly, K., Rosa, P. (1998). Transformation of the Lyme Disease Spirochete Borrelia burgdorferi with Heterologous DNA. J. Bacteriol. 180: 4850-4855 [Abstract] [Full Text]  
  • Stevenson, B., Bono, J. L., Schwan, T. G., Rosa, P. (1998). Borrelia burgdorferi Erp Proteins Are Immunogenic in Mammals Infected by Tick Bite, and Their Synthesis Is Inducible in Cultured Bacteria. Infect. Immun. 66: 2648-2654 [Abstract] [Full Text]  
  • Ge, Y., Li, C., Corum, L., Slaughter, C. A., Charon, N. W. (1998). Structure and Expression of the FlaA Periplasmic Flagellar Protein of Borrelia burgdorferi. J. Bacteriol. 180: 2418-2425 [Abstract] [Full Text]  
  • Motaleb, M. A., Corum, L., Bono, J. L., Elias, A. F., Rosa, P., Samuels, D. S., Charon, N. W. (2000). Borrelia burgdorferi periplasmic flagella have both skeletal and motility functions. Proc. Natl. Acad. Sci. USA 97: 10899-10904 [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»