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Infection and Immunity, September 2004, p. 5419-5432, Vol. 72, No. 9
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.9.5419-5432.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Combined Effects of Blood and Temperature Shift on Borrelia burgdorferi Gene Expression as Determined by Whole Genome DNA Array

Rafal Tokarz, Julie M. Anderton, Laura I. Katona, and Jorge L. Benach*

Department of Molecular Genetics and Microbiology, Center for Infectious Diseases, Stony Brook University, Stony Brook, New York

Received 14 April 2004/ Returned for modification 7 June 2004/ Accepted 22 June 2004

Borrelia burgdorferi undergoes differential gene expression during transmission from its tick vector to a vertebrate host. The addition of blood to a spirochete culture at 35°C for 48 h had a dramatic effect on gene expression of this organism. Utilizing B. burgdorferi whole genome DNA arrays, we compared the transcriptomes of the spirochetes following a 2-day temperature shift with blood and without blood. Using combined data from three independent RNA isolations we demonstrated that the addition of blood led to a differential expression of 154 genes. Of these, 75 genes were upregulated, with 49 (65%) of them encoded on plasmids. Blood supplementation of cultures also resulted in the downregulation of 79 genes, where 56 (70%) were plasmid encoded. We verified our results by reverse transcriptase PCR of several genes in both flat and feeding ticks. In the 2-day experiment we observed the effect that exposure to increased temperature and blood combined had on B. burgdorferi gene expression at this crucial time when the spirochetes begin to move from the vector to a new vertebrate host. These changes, among others, coincide with the upregulation of the chemotaxis and sensing regulons, of the lp38-encoded ABC transporter, of proteases capable of remodeling the outer surface of the spirochetes, and of the recombination genes of cp32 as a transient or initial part of the stress response of the phage. These are all functions that could cause or facilitate the changes that spirochetes undergo following a blood meal in the tick.

* Corresponding author: Mailing address: Center for Infectious Diseases, Stony Brook University, 248 Centers for Molecular Medicine, Stony Brook, NY 11794-5120. Phone: (631) 632-4286. Fax: (631) 632-4294. E-mail: jbenach{at}notes.cc.sunysb.edu.

Editor: D. L. Burns

Infection and Immunity, September 2004, p. 5419-5432, Vol. 72, No. 9
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.9.5419-5432.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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