Abstract
The budding yeast Saccharomyces cerevisiae is a useful system to express recombinant proteins and analyze protein-protein interaction. Membrane-spanning proteins like plant receptor kinases find their way to the plasma membrane when expressed in yeast and seem to retain their structure and function. Here, we describe a general yeast DNA transformation procedure based on lithium acetate, salmon sperm DNA, and polyethylene glycol used to express recombinant proteins. Yeast cells expressing plant receptor kinases can be used for in vivo and in vitro studies of receptor function.
Keywords:
Receptor kinase; Saccharomyces cerevisiae; Transformation; Yeast.
MeSH terms
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Acetates / pharmacology
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Blotting, Western / methods
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Cloning, Molecular / methods*
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DNA, Single-Stranded / genetics
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DNA, Single-Stranded / metabolism
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Electrophoresis, Polyacrylamide Gel / methods
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Galactokinase / genetics
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Galactokinase / metabolism
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Gene Expression / drug effects
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism*
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Isoenzymes / biosynthesis
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Isoenzymes / genetics
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Open Reading Frames
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Polyethylene Glycols / pharmacology
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Promoter Regions, Genetic
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Protein Domains
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Protein Kinase C / biosynthesis
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Protein Kinase C / genetics*
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Solanum lycopersicum / chemistry*
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Solanum lycopersicum / enzymology
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Solanum lycopersicum / genetics
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Transformation, Genetic
Substances
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Acetates
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DNA, Single-Stranded
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Isoenzymes
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Polyethylene Glycols
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protein kinase N
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GAL1 protein, S cerevisiae
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Galactokinase
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Protein Kinase C
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lithium acetate