Work with the Lyme Disease spirochete causative agent (Borrelia burgdorferi) and closely related strains
EMSL Project ID
3240
Abstract
The proposed work includes analyzing the global lysate of three distinct strains of B. burgdorferi to determine specific protein virulence factors. Experiments on concentrated cell samples of each strain provided by our collaborator (Xiaohua Yang, The State University of New York, Stony Brook, NY) will be performed in a biological safety level 2 (BSL2) biosafety cabinet. Cell samples will be disrupted followed by denaturization and reduction of the global protein lysate. A tryptic digestion will be performed on the lysate after which the sample will be cleaned for LCQ analysis using C18 solid phase purification. The samples will be analyzed by LCQ MS/MS to identify peptides present and results will be compared between the three strains. The three strains of interest (B31, N40, and JD1) differ dramatically in their disease causing characteristics. Infection with strain B31 causes the typical systematic infection termed Lyme disease, but strains N40 and JD1, in contrast, seem to cause only a localized skin erythema at the site of the tick bite. The entire genome of B. burgdorferi strain B31 has been determined (TIGR database), which will facilitate the characterization of proteins present in the sample. Draft sequences from the two additional B. burgdorferi strains, N40 and JD1 are in the process of being completed. It is anticipated that variations in the proteins and protein expression levels between the various strains will be detected, which will help in elucidating the virulence factors specific for each strain.
Project Details
Project type
Exploratory Research
Start Date
2003-01-16
End Date
2003-07-21
Status
Closed
Released Data Link
Team
Principal Investigator
Related Publications
Proteomic analysis of Lyme disease: Global protein comparison of three strains of Borrelia burgdorferi