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Visualization of differential gene expression by improved cyan fluorescent protein and yellow fluorescent protein production in Bacillus subtilis (2004)
| Content Provider | CiteSeerX |
|---|---|
| Author | Veening, Jan-Willem Smits, Wiep Klaas Hamoen, Leendert W. Jongbloed, Jan D. H. |
| Abstract | The distinguishable cyan and yellow fluorescent proteins (CFP and YFP) enable the simultaneous in vivo visualization of different promoter activities. Here, we report new cloning vectors for the construction of cfp and yfp fusions in Bacillus subtilis. By extending the N-terminal portions of previously described CFP and YFP variants, 20- to 70-fold-improved fluorescent-protein production was achieved. Probably, the addition of sequences encoding the first eight amino acids of the N-terminal part of ComGA of B. subtilis overcomes the slow translation initiation that is provoked by the eukaryotic codon bias present in the original cfp and yfp genes. Using these new vectors, we demonstrate that, within an isogenic population of sporulating B. subtilis cells, expression of the abrB and spoIIA genes is distinct in individual cells. The use of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria has proven to be a powerful method to study in vivo gene expression in a broad range of hosts (2). Mutagenesis of gfp resulted in variants with different fluorescent properties (4, 14). The use of the distinguishable cyan (cfp) and yellow (yfp) variants of gfp has allowed studies of multiple cellular processes within a single cell (6, 8). One of the |
| File Format | |
| Journal | Appl. Environ |
| Language | English |
| Publisher Date | 2004-01-01 |
| Access Restriction | Open |
| Subject Keyword | Bacillus Subtilis Improved Cyan Fluorescent Protein Yellow Fluorescent Protein Production Differential Gene Expression Distinguishable Cyan Broad Range Yellow Fluorescent Protein Amino Acid Yfp Variant 70-fold-improved Fluorescent-protein Production Vivo Visualization Vivo Gene Expression Single Cell Multiple Cellular Process Yfp Gene Green Fluorescent Protein Eukaryotic Codon Bias Present Different Fluorescent Property New Vector N-terminal Part N-terminal Portion Different Promoter Activity Isogenic Population Original Cfp Individual Cell Slow Translation Initiation Subtilis Cell Yfp Fusion Powerful Method Jellyfish Aequorea Victoria Spoiia Gene |
| Content Type | Text |
| Resource Type | Article |
