This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. CFP and YFP are currently the best set of fluorophores for live-cell dual imaging in S. cerevisiae. The microscopy group previously constructed plasmids carrying CFP and YFP ORFs that express well in yeast. These plasmids allow fusion of CFP or YFP to the C-terminus of any yeast protein by PCR-based integration. We altered a plasmid designed by Prein et al. so that YFP or CFP can be fused to the N-terminus of proteins by PCR-based integration. We have designed vectors to tag proteins with citrine or cerulean. We have inserted dsRED T1, Cherry and Strawberry into the integrating plasmids so that they can now be fused to the C-terminus of proteins by PCR- based integration. The center fulfilled 84 requests submitted to the Microscopy group in the last reporting year.
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