Analysis of the Development of a Morphological Phenotype as a Function of Protein Concentration in Budding Yeast

Debarati Mukherjee; Arpita Sen; R. Claudio Aguilar

doi:10.3791/1863

Analysis of the Development of a Morphological Phenotype as a Function of Protein Concentration i...

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Analysis of the Development of a Morphological Phenotype as a Function of Protein Concentration in Budding Yeast

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15:02 min

March 24, 2010

DOI: 10.3791/1863-v

Debarati Mukherjee¹, Arpita Sen¹, R. Claudio Aguilar¹

¹Department of Biological Sciences and Purdue Center for Cancer Research,Purdue University

Gene deletion and protein overexpression are common methods for studying functions of proteins. In this article, we describe a protocol for analysis of phenotype development as a function of protein concentration at population and single-cell levels in Saccharomyces cerevisiae.

This protocol utilizes a regulatable promoter that allows controlled protein expression. In this experiment, we use a high copy two micron plasmid that expresses the N two domain from a methionine repressible promoter. Wild type yeast cells are transformed with this.

Plasmid and transformants are selected on plates lacking uol. After overnight growth. In selective media cells are synchronized by no cortisol mediated growth.Arrest.

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