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Comparative Genomics of Ricketttsia prowazekii Madrid E and Breinl Strains
NAVAL MEDICAL RESEARCH CENTER SILVER SPRING MD
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Rickettsia prowazekii, the causative agent of epidemic typhus, has been responsible for millions of human deaths. Madrid E is attenuated strain of R. prowazekii, while Breinl is a virulent strain. The genomic DNA sequence of Madrid E has recently been published. To study the genomic variations between Madrid E reference and Breinl test DNAs, cohybridization experiments were performed on a DNA microarray containing all 834 protein-coding genes of Madrid E. Of the 834 genes assessed, 24 genes showed 1.5-to 2.0-fold increases in hybridization signals in Breinl DNA compared to Madrid E DNA, Indicating the presence of genomic variations in 3 of the total genes. Eighteen of these 24 genes are predicted to be involved in different functions. Southern blot analysis of five genes, virB4, ftsK, rfbE, lpxA, and rpoH, suggested the presence of an additional paralogs in Breinl, which might be related to the observed increase in hybridization signals. Studies by real-time reverse transcription-PCR revealed an increase in expression of the above-mentioned five genes and five other genes. In addition to the elevated hybridization signals of 24 genes observed in the Breinl strain, one gene rp084 showed only 110 the hybridization signal of Madrid E. Further analysis of this gene by PCR and sequencing revealed a large deletion flanking the whole rp084 gene and part of the rp083 gene in the virulent Breinl strain. The results of this first rickettsial DNA microarray may provide some important information for the elucidation of pathogenic mechanisms of R. prowazekii.
APPROVED FOR PUBLIC RELEASE