Using genomic fingerprinting by arbitrarily primed PCR (AP-PCR), we have shown that the species Borrelia burgdorferi is actually comprised of at least three distinct phyletic groups, whose level of divergence approaches that of distinct species by several criteria (Welsh et al., 1992). The validity of AP-PCR as a phylogenetic tool has been corroborated by multilocus enzyme electrophoresis (MEE) and other methods, as we will demonstrate. Here, we propose to apply genomic fingerprinting and other methods to resolve several questions in the population genetics of B. burgdorferi. First, we will compare phylogenetic analysis using AP-PCR with MEE for the ECOR collection. We will refine the correlation between genetic distances measured by MEE and AP-PCR using this data. Second, we will continue to explore the number and distribution of distinct phyletic groups of B. burgdorferi, expecting to reveal distinct species or subspecies, and generally outline the genetic structure of the population. In this part of the study, we will determine whether the intraspecies phylogeny of the spirochete correlates with regional geographics within North America, as has been demonstrated for B. burgdorferi in Eurasia. We will also compare Group I B. burgdorferi from both continents, to determine the genetic relationship between the two populations. Third, we will determine if B. burgdorferi from different phyletic groups have different preferred hosts, including human. Fourth, we will seek evidence of naturally occurring genetic recombination between strains of B. burgdorferi by comparing DNA sequence information and constructing gene trees. Fifth, we will seek evidence for the horizontal movement of plasmids between different branches of the phylogenetic tree, using AP-PCR homoplasy, comparisons of plasmid profiles and Southern blotting.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29AI032644-01A1
Application #
3456121
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1992-12-01
Project End
1997-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
California Institute of Biological Research
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Dalal, S S; Welsh, J; Tkachenko, A et al. (1994) Rapid isolation of tissue-specific and developmentally regulated brain cDNAs using RNA arbitrarily primed PCR (RAP-PCR). J Mol Neurosci 5:93-104
McClelland, M; Welsh, J (1994) RNA fingerprinting by arbitrarily primed PCR. PCR Methods Appl 4:S66-81
Wong, K K; McClelland, M (1994) Stress-inducible gene of Salmonella typhimurium identified by arbitrarily primed PCR of RNA. Proc Natl Acad Sci U S A 91:639-43
Wong, K K; Wong, R M; Rudd, K E et al. (1994) High-resolution restriction map for a 240-kilobase region spanning 91 to 96 minutes on the Salmonella typhimurium LT2 chromosome. J Bacteriol 176:5729-34
Ralph, D; Postic, D; Baranton, G et al. (1993) Species of Borrelia distinguished by restriction site polymorphisms in 16S rRNA genes. FEMS Microbiol Lett 111:239-43
Ralph, D; Que, Q; Van Etten, J L et al. (1993) Leptospira genomes are modified at 5'-GTAC. J Bacteriol 175:3913-5
Ralph, D; McClelland, M; Welsh, J (1993) RNA fingerprinting using arbitrarily primed PCR identifies differentially regulated RNAs in mink lung (Mv1Lu) cells growth arrested by transforming growth factor beta 1. Proc Natl Acad Sci U S A 90:10710-4
Ralph, D; McClelland, M (1993) Intervening sequence with conserved open reading frame in eubacterial 23S rRNA genes. Proc Natl Acad Sci U S A 90:6864-8
Ralph, D; McClelland, M; Welsh, J et al. (1993) Leptospira species categorized by arbitrarily primed polymerase chain reaction (PCR) and by mapped restriction polymorphisms in PCR-amplified rRNA genes. J Bacteriol 175:973-81
Welsh, J; Chada, K; Dalal, S S et al. (1992) Arbitrarily primed PCR fingerprinting of RNA. Nucleic Acids Res 20:4965-70