Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4870

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Imaging Real-Time Gene Expression in Living Yeast

Daniel Zenklusen1, Amber L Wells1, John S. Condeelis1,2, and Robert H. Singer1,2,3

1 Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA
2 Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA

3Corresponding author (rhsinger{at}aecom.yu.edu)


INTRODUCTION

This protocol describes the application of the MS2 system to the yeast Saccharomyces cerevisiae. ASH1 mRNA tagged with six MS2 repeats (6MBSs) is used to follow the localization of the ASH1 mRNA particles to the bud tip of a haploid yeast cell. W303 yeast cells transformed with pG14-MS2-GFP and pGAL-lacZ-MS2-ASH1 are grown on select medium lacking tryptophan and leucine. RNA expression is induced by the addition of galactose, and a time-lapse movie is then acquired.


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