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PMID: 17417645 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A genomic integration method to visualize localization of endogenous mRNAs in living yeast.

Nature methods ·Vol. 4 ·No. 5 ·2007-05-00 ·Pages 409-12

Haim L, Zipor G, Aronov S, Gerst JE

Abstract

mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2-CP) between the coding region and 3' untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae).

MeSH Terms
Adaptor Proteins, Signal Transducing Capsid Proteins/genetics Carrier Proteins/metabolism DNA-Binding Proteins/metabolism Green Fluorescent Proteins/chemistry Levivirus/genetics Membrane Proteins/metabolism Microscopy, Fluorescence Peroxins Polymerase Chain Reaction/methods RNA, Fungal/analysis,metabolism RNA, Messenger/analysis,metabolism RNA-Binding Proteins/genetics Repressor Proteins/metabolism Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins/metabolism Schizosaccharomyces/genetics
Chemicals
ASH1 protein, S cerevisiae Adaptor Proteins, Signal Transducing Capsid Proteins Carrier Proteins DNA-Binding Proteins Membrane Proteins PEX3 protein, S cerevisiae Peroxins RNA, Fungal RNA, Messenger RNA-Binding Proteins Repressor Proteins SRO7 protein, S cerevisiae Saccharomyces cerevisiae Proteins Green Fluorescent Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Haim Liora
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
Zipor Gadi
Aronov Stella
Gerst Jeffrey E
Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7091
DOI
10.1038/nmeth1040
Published
2007-05-00
Epub
2007-00-08
Pages
409-12
Language
English
Region
United States
NLM ID
101215604
Subset
IM
Corrections
CommentIn
PMID: 17464293
External Links
PubMed source DOI: 10.1038/nmeth1040 View journal details
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