A conserved, transcriptionally active p28 multigene locus of Ehrlichia canis

Antigenic diversity of Ehrlichia chaffeensis and Ehrlichia canis may involve independent or differential expression of the P28 outer membrane proteins genes, enabling persistent infections of the natural hosts. In this study, we analyzed the transcriptional activity of a five gene locus in E. canis encoding homologous, but non-identical, p28 genes. The p28 multigene locus contained three previously identified complete gene sequences and one partial gene sequence. A new p28 gene was identified and sequenced, and the complete sequence of a second partial p28 gene was determined. The new p28 gene joined two previously separate loci, forming the single p28 multigene locus. The amino acid homology of the E. canis P28 proteins ranged from 51 to 74%. The nucleic acid sequence of regions compared within the locus spanning four p28 genes from two geographically distinct E. canis isolates was completely conserved. Analysis of the five p28 genes demonstrated that all were transcriptionally active in in-vitro cultures of E. canis incubated at the vertebrate host (37 degrees C) and ambient tick temperatures (27 degrees C). Polycistronic copies of multiple p28 genes were not detected by RT-PCR, but monocistronic p28 mRNA transcripts were detected by Northern blotting from E. canis infected DH82 cells, indicating that the genes are transcribed as monocistronic messages.