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Information × Registration Number 0217U000773, 0112U003243 , R & D reports Title Yeast cell as a model for the study of signaling pathways involved in the development of pathological processes in the human body popup.stage_title Head Sybirna Nataliia Olexandrivna, Stasyk Olena Georgiivna, Registration Date 28-02-2017 Organization Ivan Franko National University of Lviv popup.description2 As a result of this work vector for expression of the chimeric genetic constructs yEGFP3-SNCA, encoding human alpha-synuclein fused to green fluorescent protein (EGFP) was constructed, and the model H. polymorpha yeast strains were obtained. The presence of the heterologous protein alpha-synuclein conjugated with EGFP in the cells of model yeast strains was detected by fluorescent microscopy and Western blot analysis. To simulate aging of neurons at Parkinson's disease, which is inherent in glucose hypometabolism, strains producing alpha-synuclein person with impaired glucose transport were constructed. It has been found that the production and accumulation of human alpha-synuclein in methylotrophic yeast H. polymorpha cells caused damage in the growth and adversely affects the cell viability. Deficiency of glucose as the carbon source had a negative impact on model strains, as well as enhanced the cytotoxicity of alpha-synuclein, whereas an excess of glucose in the cultural medium increased the resistance of the model yeast cells to production of heterologous strains of alpha-synuclein. The marker of arginine auxotrophy (deletion of HpASS1gene encoding the enzyme arginine succinate synthase) has also been introduced in model yeast strains. It made possible to create conditions for yeast cells, similar to those that arise as a result of the limitation of the human body for this semi-essential amino acid. By creating a limit of exogenous arginine for ass1 yeast mutant, we can induce and regulate autophagy process in yeast cells and to explore the processes of degradation of alpha-synuclein aggregates. Product Description popup.authors Денега Ірина Ігорівна Климишин Наталія Ігорівна popup.nrat_date 2020-04-02 Close