Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging

Wout Overkamp, Katrin Beilharz, Ruud Detert Oude Weme, Ana Solopova, Harma Karsens, Ákos T. Kovács, Jan Kok, Oscar P. Kuipers, Jan Willem Veening*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review


Green fluorescent protein (GFP) offers efficient ways of visualizing promoter activity and protein localization in vivo, and many different variants are currently available to study bacterial cell biology. Which of these variants is best suited for a certain bacterial strain, goal, or experimental condition is not clear. Here, we have designed and constructed two "superfolder" GFPs with codon adaptation specifically for Bacillus subtilis and Streptococcus pneumoniae and have benchmarked them against five other previously available variants of GFP in B. subtilis, S. pneumoniae, and Lactococcus lactis, using promoter-gfp fusions. Surprisingly, the best-performing GFP under our experimental conditions in B. subtilis was the one codon optimized for S. pneumoniae and vice versa. The data and tools described in this study will be useful for cell biology studies in low-GC-rich Gram-positive bacteria.

Original languageEnglish
JournalApplied and Environmental Microbiology
Issue number20
Pages (from-to)6481-6490
Publication statusPublished - 2013
Externally publishedYes

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