Identification of Eimeria bovis merozoite cDNAs using differential mRNA display

Abstract

Differences in gene expression between Eimeria bovis sporozoites and first-generation merozoites were analyzed using the technique of differential mRNA display. Approx. 5% of the sequences detected in first-generation merozoites appear to be unique relative to sporozoites. Several of the bands corresponding to merozoite-specific gene expression were isolated and cloned. Northern blot analysis revealed that the cDNA fragments DMZ-7, DMZ-8 and NMZ-6 hybridized to mRNAs expressed at > 50-fold higher levels in merozoites relative to sporozoites. A fourth cDNA fragment, NMZ-4, hybridized to a mRNA expressed at 3-fold higher levels in merozoites. Further characterization demonstrated that expression of DMZ-8 in E. bovis-infected bovine cells begins as early as 12 h after sporozoite invasion and continues throughout the entire 14 days of first-generation schizogony. Sequence analysis of each of the four merozoite cDNAs failed to identify any significant similarity to any entries in the GenBank database, suggesting that these developmentally regulated genes may be unique to coccidian parasites.