title: Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging.
abstract: 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.
authors: Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, Centre for Synthetic Biology, University of Groningen, Groningen, The Netherlands.
authors: Wout
authors: W
authors: Overkamp
authors: Katrin
authors: K
authors: Beilharz
authors: Ruud
authors: R
authors: Detert Oude Weme
authors: Ana
authors: A
authors: Solopova
authors: Harma
authors: H
authors: Karsens
authors: Ákos  T
authors: Á
authors: Kovács
authors: Jan
authors: J
authors: Kok
authors: Oscar P
authors: OP
authors: Kuipers
authors: Jan-Willem
authors: JW
authors: Veening
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019985:Benchmarking; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D013294:Lactococcus lactis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling / Q000379:methods*; D013296:Streptococcus pneumoniae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D049452:Green Fluorescent Proteins
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