Isolation of a Toxoplasma gondii cyclin by yeast two-hybrid interactive screen.

摘要: GAL4-based yeast two-hybrid cDNA libraries from Toxoplasma gondii RH strain were constructed and screened for interactors of a putative T. cdc2-related kinase, TgCRK2. A screen 3.2 million transformants yielded single clone that harbored protein fusion capable specifically interacting with Sequencing revealed the insert (TgCYC1) had homology to cyclin class proteins. The TgCYC1 fragment was used probe conventional library 2.65 kb coding predicted 582 amino acids obtained. Based on comparison 5'-RACE product tachyzoite mRNA, appeared be complete. highest similarity Plasmodium falciparum CYC1 displayed sequence characteristics place it in H eukaryotic cyclins. In synchronous populations level mRNA unchanged indicating is not cell cycle regulated at level. rescues G(1)/S defect S. cerevisiae DL1 demonstrating this apicomplexan can function an established heterologous model system.