Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 79, Issue 20, Pages 6481-6490Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.02033-13
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Funding
- STW [107504]
- ALW [81807004]
- Kluyver Centre for Genomics of Industrial Fermentation
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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.
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