The calmodulin-ubiquitin (CUB) genes of Trypanosoma cruzi are essential for parasite viability

The CUB genes represent single copy genes in the diploid Trypanosoma cruzi genome. In this report data are presented which demonstrate that a single expressed CUB gene is necessary for parasite viability. Although either CUB gene could be deleted individually, repeated attempts to simultaneously delete both genes were unsuccessful. The essential nature of the CUB genes was further supported by studies which demonstrated positive selection for CUB gene expression. Positive selection was demonstrated by carrying out dual gene replacements which showed that both native CUB genes could be efficiently deleted provided the CalB1 calmodulin gene was simultaneously replaced by a CUB gene protein coding sequence. Although the function of the CUB gene product remains unknown the experiments presented here indicate the product is likely to play an important role in the parasites' life cycle.