RRC ID 48076
著者 Kaishima M, Ishii J, Matsuno T, Fukuda N, Kondo A.
タイトル Expression of varied GFPs in Saccharomyces cerevisiae: codon optimization yields stronger than expected expression and fluorescence intensity.
ジャーナル Sci Rep
Abstract Green fluorescent protein (GFP), which was originally isolated from jellyfish, is a widely used tool in biological research, and homologs from other organisms are available. However, researchers must determine which GFP is the most suitable for a specific host. Here, we expressed GFPs from several sources in codon-optimized and non-codon-optimized forms in the yeast Saccharomyces cerevisiae, which represents an ideal eukaryotic model. Surprisingly, codon-optimized mWasabi and mNeonGreen, which are typically the brightest GFPs, emitted less green fluorescence than did the other five codon-optimized GFPs tested in S. cerevisiae. Further, commercially available GFPs that have been optimized for mammalian codon usage (e.g., EGFP, AcGFP1 and TagGFP2) unexpectedly exhibited extremely low expression levels in S. cerevisiae. In contrast, codon-optimization of the GFPs for S. cerevisiae markedly increased their expression levels, and the fluorescence intensity of the cells increased by a maximum of 101-fold. Among the tested GFPs, the codon-optimized monomeric mUkG1 from soft coral showed the highest levels of both expression and fluorescence. Finally, the expression of this protein as a fusion-tagged protein successfully improved the reporting system's ability to sense signal transduction and protein-protein interactions in S. cerevisiae and increased the detection rates of target cells using flow cytometry.
巻・号 6
ページ 35932
公開日 2016-10-26
DOI 10.1038/srep35932
PII srep35932
PMID 27782154
PMC PMC5080575
MeSH Animals Anthozoa / genetics Anthozoa / metabolism Codon / genetics Flow Cytometry Gene Expression Green Fluorescent Proteins / genetics* Green Fluorescent Proteins / metabolism* Luminescent Proteins / genetics Luminescent Proteins / metabolism Microscopy, Fluorescence Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Recombinant Proteins / genetics Recombinant Proteins / metabolism Saccharomyces cerevisiae / genetics* Saccharomyces cerevisiae / metabolism* Sequence Tagged Sites Signal Transduction
IF 3.998
引用数 16
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
酵母 pGK416