Isolation and characterization of RNA from the intracellular parasite Theileria parva

Abstract

Using a rapid procedure to isolate schizonts of the intracellular parasite Theileria parva from infected bovine lymphocytes, we have prepared parasite RNA that is more than 90% pure. Characterization of this schizont RNA has revealed the presence of two ribosomal RNA species of 3.3 kb and 1.8 kb and a third non-adenylated abundant RNA species of 1.9 kb. In vitro translation of the isolated schizont mRNA has identified about 200 parasite specific polypeptides, only a few of which could be detected by translation of mRNA from infected host cells. By analysing the kinetics of liquid hybridization of schizont mRNA with its homologous complementary DNA the nucleotide sequence complexity of the abundant class of the parasite mRNA has been estimated to be 1.7 × 10 3 kb. Assuming a number average size of 2 kb per mRNA molecule this would represent 4000 transcripts for all abundance classes of the schizont mRNA. Using the same technique we estimate that approximately 10% of the mRNA isolated from infected lymphocytes were transcripts from the parasite genome. We conclude that the low number of parasite specific translation products in the mRNA from infected lymphocytes and the low number of parasite proteins detected in isolated schizonts reported previously is due to the low abundance of the parasite transcripts rather than a low number of expressed parasite genes.