Expression of fluorescent genes in Trypanosoma cruzi and Trypanosoma rangeli (Kinetoplastida: Trypanosomatidae): its application to parasite-vector biology.
We describe here the identification of the calmodulin-ubiquitin associated (CUB) genes of Trypanosoma cruzi. A single CUB gene resides in a 1.5-kb DNA sequence linking the calmodulin and ubiquitin genes in the 2.65 and 2.8 loci (CUB2.65 and CUB2.8 respectively). The CUB genes also share the same coding strand as the flanking calmodulin and ubiquitin genes. DNA sequence analysis reveals that each CUB gene contains an open reading frame which would encode a protein of 208 amino acids. The CUB protein shares homology with the recently identified calcium binding EFH5 protein of T. brucei. Transcription of the CUB genes results in the generation of a mRNA of approximately 1.0 kb. CUB cDNA sequence analysis following PCR amplification of the CUB mRNA population indicates that both genes are expressed and trans-spliced, but utilize different 3' acceptor sites for the trans-splicing reaction.