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- W2979227170 abstract "Yeast are an ideal system to study Heat Shock Protein 70 (Hsp70) function in a cellular context. This protocol was generated to analyze the function of non-native Hsp70 proteins by expressing them as the sole cytosolic Hsp70 in yeast. As an initial step, Hsp70 variants (such as Ssa1 point mutants and non-yeast versions such as Nematostella vectensis NvHsp70A, B and D) are cloned into an appropriate expression plasmid. Next, these plasmids are transformed into ssa1-4Δ yeast [expressing native Ssa1 from an uracil-based (URA3) plasmid] which are subsequently cured of the original yeast on 5-Fluroorotic Acid (5-FOA). The resulting cells can be screened for a variety of phenotypes which match to the activity of well-studied cellular pathways." @default.
- W2979227170 created "2019-10-10" @default.
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- W2979227170 date "2019-01-01" @default.
- W2979227170 modified "2023-09-23" @default.
- W2979227170 title "Analyzing the Functionality of Non-native Hsp70 Proteins in Saccharomyces cerevisiae" @default.
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- W2979227170 doi "https://doi.org/10.21769/bioprotoc.3389" @default.
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