Borrelia burgdorferi is an in vitro culturable bacterium that is the cause of Lyme disease. Its small genome contain approxnnately 1,000 chromosomal genes and 400 plasmid genes. Despite knowledge of the complete DNA sequence of the B. burgdorferi genome, identification and characterization of unique in vivo expressed B. burgdorferi virulence determinants has been delayed by the lack of expeditious and efficacious genetic systems in Borrelia. We have developed a genetic system in B. burgdorfed that consists of the extrachromosomal cloning vector, pGKI2, enhanced green fluorescent protein as a potential reporter gene, and resistance to erythromycin, kanamycin, and other antibiotics as selective markers. We have also been able to show that the bmp gene cluster is highly conserved among B. burgdorferi sensu lato strains, and that the genes of this cluster undergo environmentally modulated differential expression suggesting a potential role in virulence for these genes. The experimental protocol we propose is based on our preliminary work and framed by two hypotheses: 1) efficient molecular genetic systems can be developed for B. burgdorferi, and 2) the role of the bmp gene cluster in B.burgdorferi biology and virulence can be ascertained using these systems. With the long-term aim of identifying B.burgdorferi virulence determinants and improving our understanding of their in vivo expression and regulation, we propose the following Specific Aims: 1) Continue development and improvement of an extrachromosomal cloning system for B. burgdorferi, 2) isolate and complement B. burgdorferi bmpD, bmpC and bmpA null mutants to determine the possible role of these genes in B. burgdorferi virulence in in vitro and in vivo model systems of infection; and 3) characterize promoters and regulatory DNA sequences of bmpC using transcriptional fusions with enhanced green fluorescent protein (EGFP) and fluorescence-activated cell sorting (FACS). We expect these experiments will permit the extension of the molecular Koch's postulates to the characterization of unique aud specific molecular virulence determinants of B. burgdorferi.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI048856-05
Application #
6844365
Study Section
Special Emphasis Panel (ZRG1-MBC-1 (01))
Program Officer
Baker, Phillip J
Project Start
2001-02-01
Project End
2007-01-31
Budget Start
2005-02-01
Budget End
2007-01-31
Support Year
5
Fiscal Year
2005
Total Cost
$380,901
Indirect Cost
Name
New York Medical College
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041907486
City
Valhalla
State
NY
Country
United States
Zip Code
10595
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Huang, Weihua; Ojaimi, Caroline; Fallon, John T et al. (2015) Genome Sequence of Borrelia chilensis VA1, a South American Member of the Lyme Borreliosis Group. Genome Announc 3:
Bugrysheva, Julia V; Pappas, Christopher J; Terekhova, Darya A et al. (2015) Characterization of the RelBbu Regulon in Borrelia burgdorferi Reveals Modulation of Glycerol Metabolism by (p)ppGpp. PLoS One 10:e0118063
Ivanova, Larisa B; Tomova, Alexandra; González-Acuña, Daniel et al. (2014) Borrelia chilensis, a new member of the Borrelia burgdorferi sensu lato complex that extends the range of this genospecies in the Southern Hemisphere. Environ Microbiol 16:1069-80
Sambir, Mariya; Ivanova, Larisa B; Bryksin, Anton V et al. (2011) Functional analysis of Borrelia burgdorferi uvrA in DNA damage protection. FEMS Microbiol Lett 317:172-80
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Bryksin, Anton V; Tomova, Alexandra; Godfrey, Henry P et al. (2010) BmpA is a surface-exposed outer-membrane protein of Borrelia burgdorferi. FEMS Microbiol Lett 309:77-83
Cabello, Felipe C; Godfrey, Henry P; Newman, Stuart A (2007) Hidden in plain sight: Borrelia burgdorferi and the extracellular matrix. Trends Microbiol 15:350-4
Moreno, Claudia X; Moy, Fred; Daniels, Thomas J et al. (2006) Molecular analysis of microbial communities identified in different developmental stages of Ixodes scapularis ticks from Westchester and Dutchess Counties, New York. Environ Microbiol 8:761-72
Shin, Junghee J; Bryksin, Anton V; Godfrey, Henry P et al. (2004) Localization of BmpA on the exposed outer membrane of Borrelia burgdorferi by monospecific anti-recombinant BmpA rabbit antibodies. Infect Immun 72:2280-7

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