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Infection and Immunity, June 2001, p. 3536-3541, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.3536-3541.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Purification and Characterization of Borrelia burgdorferi from Feeding Nymphal Ticks (Ixodes scapularis)

Sivaprakash Rathinavelu and Aravinda M. de Silva*

Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina 27599

Received 22 January 2001/Returned for modification 22 February 2001/Accepted 8 March 2001

Here we describe a protocol for purifying Borrelia burgdorferi from feeding ticks by velocity centrifugation and Percoll density gradient centrifugation. The purified spirochetes were motile and 10- to 20-fold purer than the bacteria in crude tick homogenates. The purified bacteria were present in sufficient quantity for protein and gene expression studies. In comparison to culture-grown bacteria, tick-borne spirochetes had several proteins that were upregulated and a few that were downregulated. When the levels of B. burgdorferi outer surface proteins OspA and OspC were measured, OspC protein and mRNA levels were lower in cultured bacteria than in bacteria purified from ticks. Although differences in OspA mRNA levels were observed between cultured and tick-borne bacteria, no differences were observed at the protein level. These experiments demonstrate that tick-transmitted borreliae display a gene expression and antigen profile different from that of spirochetes cultured in vitro.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, CB#7290 University of North Carolina, Chapel Hill, NC 27599. Phone: (919) 843-9964. Fax: (919) 962-8103. E-mail: desilva{at}med.unc.edu.

Infection and Immunity, June 2001, p. 3536-3541, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.3536-3541.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • von Lackum, K., Miller, J. C., Bykowski, T., Riley, S. P., Woodman, M. E., Brade, V., Kraiczy, P., Stevenson, B., Wallich, R. (2005). Borrelia burgdorferi Regulates Expression of Complement Regulator-Acquiring Surface Protein 1 during the Mammal-Tick Infection Cycle. Infect. Immun. 73: 7398-7405 [Abstract] [Full Text]  
  • Miller, J. C., von Lackum, K., Babb, K., McAlister, J. D., Stevenson, B. (2003). Temporal Analysis of Borrelia burgdorferi Erp Protein Expression throughout the Mammal-Tick Infectious Cycle. Infect. Immun. 71: 6943-6952 [Abstract] [Full Text]  
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