一种用于可视化活酵母中内源性mRNA定位的基因组整合方法。
A genomic integration method to visualize localization of endogenous mRNAs in living yeast.
作者信息
Haim Liora, Zipor Gadi, Aronov Stella, Gerst Jeffrey E
机构信息
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
出版信息
Nat Methods. 2007 May;4(5):409-12. doi: 10.1038/nmeth1040. Epub 2007 Apr 8.
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).
信使核糖核酸（mRNA）定位可能是蛋白质定位的一个重要决定因素。我们描述了一种基于聚合酶链式反应（PCR）的简单基因组标记策略（m-TAG），该策略利用同源重组在任何酵母基因的编码区和3'非翻译区（UTR）之间插入RNA结合性MS2外壳蛋白（MS2-CP）的结合位点。在与绿色荧光蛋白（GFP）融合的MS2-CP共表达时，我们证明了内源性mRNA（ASH1、SRO7、PEX3和OXA1）在活酵母（酿酒酵母）中的定位。
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