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Information × Registration Number 0211U013054, 0110U006325 , R & D reports Title Studying the selective autophagic degradation of organelles and cytosolic proteins in yeasts Pichia pastoris and Saccharomyces cerevisiae popup.stage_title Head Sibirny Andriy Andriyovych, Registration Date 28-05-2012 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 On this stage of work it was planned to select the conditions for the investigation of pexophagy in yeast Saccharomyces cerevisiae on plates, insertional mutagenesis of wild type of this yeast specie and search for potential atg mutants. On the previous stage of work we constructed vector, containing the gene of beta-galactosidase (LacZ) with the DNA sequence that encodes the signal tripeptide SKL (Serine - Lysine-Leucine) at С-terminus for the delivery of protein into peroxisomes under the promotor of FOX3 gene. The test on plates with a minimum medium with an oleate and X-gal demonstrated that the protein is expressed on oleate and possesses enzymatic activity. On this stage of work it was shown with the help of light microscopy that hybrid protein LacZ-SKL is localized in peroxisomes on oleate medium with X-gal. The conditions for successful inactivation and degradation of this enzyme after the transfer of the strain from the oleate medium with X-gal to the glucose medium without nitrogen source and with X-gal. The insertional mutagenesis of the wild type containing PFOX3-LacZ-SKL was carried out. Under the elaborated conditions the mutants with the affected inactivation/degradation of this enzyme were selected, that indicates the defect of peroxisome degradation in these strains. Product Description popup.authors Назарко Володмир Юрійович Полупанов Андрій Сергійович Сибірний Андрій Андрійович popup.nrat_date 2020-04-02 Close