CONDITIONAL GENE SILENCING IN EUKARYOTES BY PROMOTER INSERTION

摘要

The Renilla reniformis luciferase (RLUC) gene was used as a bioluminescent reporter to investigate promoter silencing in the opportunistic fungus Candida albicans. Five expression modules, containing the RLUC) coding sequence under transcriptional control of a yeast promoter, were cloned downstream of the strong inducible GAL1 or MAL2 promoter of C. albicans. After transformation and growth under inducing conditions, luciferase activity was inhibited up to 96 % as a result of upstream transcription initiation and elongated transcript formation. Similar silencing results were obtained in P. pastoris, by placing the constitutive OCH1 promoter behind the inducible AOX1) promoter. Repression of the OCH1p after induction of the AOX1) promoter results in a change in glycosylation pattern.