| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
University of Rochester Medical Center, 601 Elmwood Ave, Box 672, Rochester, NY 14642, USA
Correspondence
Martin S. Pavelka, Jr
martin_pavelka{at}urmc.rochester.edu
This paper is the first detailed description of the development and use of new genetic tools specifically for the safe manipulation of highly pathogenic Francisella tularensis subsp. tularensis. Most of these tools are also demonstrated to work with other F. tularensis subspecies. Kanamycin and hygromycin resistance determinants that function as genetic markers in F. tularensis subsp. tularensis strain Schu and sets of episomal shuttle vectors that are either unstable or stably maintained in the absence of selection were developed. In addition, the hyg gene, expressed from the F. tularensis groESL promoter, was successfully used as a marker for transposon mutagenesis. This work also includes the development of sacB-based suicide plasmids expressing kanamycin resistance that can be used for electroporation-mediated allelic exchange of unmarked mutations in Schu and the F. tularensis live vaccine strain (LVS). Using these plasmids, the two predicted 
-lactamase genes, blaA and blaB, in Schu and LVS were deleted. Only the 
blaB1 mutants had increased susceptibility to ampicillin, and this phenotype was complemented by a plasmid expressing blaB+. The results suggest that the -lactam antibiotic resistance phenotype of Schu and LVS is likely due to only one of the two -lactamase genes present and that ampicillin resistance can be used as an additional selectable marker in -lactamase deletion mutants. The collection of tools presented in this report will be helpful for the genetic analyses of F. tularensis subsp. tularensis pathogenesis.
These authors contributed equally to this work.
Present address: Biology Department, University of Maryland, Baltimore County, Baltimore, MD 21250, USA.
This article has been cited by other articles:
|
|
 |
|
  S. Chakraborty, M. Monfett, T. M. Maier, J. L. Benach, D. W. Frank, and D. G. Thanassi Type IV Pili in Francisella tularensis: Roles of pilF and pilT in Fiber Assembly, Host Cell Adherence, and Virulence Infect. Immun., July 1, 2008; 76(7): 2852 - 2861. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. S. Ludu, O. M. de Bruin, B. N. Duplantis, C. L. Schmerk, A. Y. Chou, K. L. Elkins, and F. E. Nano The Francisella Pathogenicity Island Protein PdpD Is Required for Full Virulence and Associates with Homologues of the Type VI Secretion System J. Bacteriol., July 1, 2008; 190(13): 4584 - 4595. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Rodriguez, J.-J. Yu, G. Davis, B. P. Arulanandam, and K. E. Klose Targeted Inactivation of Francisella tularensis Genes by Group II Introns Appl. Envir. Microbiol., May 1, 2008; 74(9): 2619 - 2626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. W. Buchan, M. K. McLendon, and B. D. Jones Identification of Differentially Regulated Francisella tularensis Genes by Use of a Newly Developed Tn5-Based Transposon Delivery System Appl. Envir. Microbiol., May 1, 2008; 74(9): 2637 - 2645. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Horzempa, D. M. Tarwacki, P. E. Carlson Jr., C. M. Robinson, and G. J. Nau Characterization and Application of a Glucose-Repressible Promoter in Francisella tularensis Appl. Envir. Microbiol., April 1, 2008; 74(7): 2161 - 2170. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Maier, M. S. Casey, R. H. Becker, C. W. Dorsey, E. M. Glass, N. Maltsev, T. C. Zahrt, and D. W. Frank Identification of Francisella tularensis Himar1-Based Transposon Mutants Defective for Replication in Macrophages Infect. Immun., November 1, 2007; 75(11): 5376 - 5389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Su, J. Yang, D. Zhao, T. H. Kawula, J. A. Banas, and J.-R. Zhang Genome-Wide Identification of Francisella tularensis Virulence Determinants Infect. Immun., June 1, 2007; 75(6): 3089 - 3101. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. W. FRANK and T. C. ZAHRT Genetics and Genetic Manipulation in Francisella Tularensis Ann. N.Y. Acad. Sci., June 1, 2007; 1105(1): 67 - 97. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. BARKER and K. E. KLOSE Molecular and Genetic Basis of Pathogenesis in Francisella Tularensis Ann. N.Y. Acad. Sci., June 1, 2007; 1105(1): 138 - 159. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. WAYNE CONLAN and P. C.F. OYSTON Vaccines Against Francisella Tularensis Ann. N.Y. Acad. Sci., June 1, 2007; 1105(1): 325 - 350. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. W. TITBALL, A. SJOSTEDT, M. S. PAVELKA JR, and F. E. NANO Biosafety and Selectable Markers Ann. N.Y. Acad. Sci., June 1, 2007; 1105(1): 405 - 417. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
