Visualization of differential gene expression by improved cyan fluorescent protein and yellow fluorescent protein production in Bacillus subtilis

Jan Willem Veening, Wiep Klaas Smits, Leendert W. Hamoen, Jan D.H. Jongbloed, Oscar P. Kuipers*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)6809-6815
Number of pages7
JournalApplied and Environmental Microbiology
Volume70
Issue number11
DOIs
Publication statusPublished - 1 Nov 2004
Externally publishedYes

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