Optimisation of the Schizosaccharomyces pombe urg1 Expression System

• DOI: 10.1371/journal.pone.0083800
• Publication Date: 2013-12-20T21:50:01Z
• Authors: Adam T. Watson; Yasukazu Daigaku; Saed Mohebi; Thomas J. Etheridge; Charly Chahwan; Johanne M. Murray; Antony M. Carr
• Keywords: Degron; Schizosaccharomyces; Transcription

The ability to study protein function in vivo often relies on systems that regulate the presence and absence of interest. Two limitations for previously described transcriptional control are used expression fission yeast are: time taken inducing conditions initiate transcription achieve very low basal "OFF-state". In previous work, we a Cre recombination-mediated system allows rapid efficient regulation any gene interest by urg1 promoter, which has dynamic range approximately 75-fold is induced within 30-60 minutes uracil addition. this report describe easy-to-use versatile modules can be exploited significantly tune down Purg1 "OFF-levels" while maintaining an equivalent range. We also provide plasmids tools combining with auxin degron tag help maintain null-like phenotype. demonstrate utility improved HO-dependent site-specific DSB formation, Rtf1-dependent replication fork arrest controlling Rhp18Rad18-dependent post repair.