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; Akos T. Kovacs; Jan Kok; Oscar P. Kuipers; Jan-Willem Veening

doi:10.1128/AEM.02033-13

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, Akos T. Kovacs, Jan Kok, Oscar P. Kuipers, Jan-Willem Veening^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

73 Citations (Scopus)

510 Downloads (Pure)

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.

Original language	English
Pages (from-to)	6481-6490
Number of pages	10
Journal	Applied and environmental microbiology
Volume	79
Issue number	20
DOIs	https://doi.org/10.1128/AEM.02033-13
Publication status	Published - Oct-2013

Keywords

GENE REGULATORY NETWORKS
QUANTITATIVE-ANALYSIS
OROTATE TRANSPORTER
CODON USAGE
EXPRESSION
COMPETENCE
BACTERIA
PLASMID
GENOME
TRANSFORMATION

Access to Document

10.1128/AEM.02033-13

2013ApplEnvironMicrobiolOverkampSupp.pdfFinal publisher's version, 328 KB
2013ApplEnvironMicrobiolOverkamp.pdfFinal publisher's version, 1.5 MB

Handle.net

Persistent link

Cite this

Overkamp, W., Beilharz, K., Weme, R. D. O., Solopova, A., Karsens, H., Kovacs, A. T., Kok, J., Kuipers, O. P., & Veening, J-W. (2013). Benchmarking Various Green Fluorescent Protein Variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for Live Cell Imaging. Applied and environmental microbiology, 79(20), 6481-6490. https://doi.org/10.1128/AEM.02033-13

@article{d4970610ab55404cb65f5b3a36be31de,

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.",

keywords = "GENE REGULATORY NETWORKS, QUANTITATIVE-ANALYSIS, OROTATE TRANSPORTER, CODON USAGE, EXPRESSION, COMPETENCE, BACTERIA, PLASMID, GENOME, TRANSFORMATION",

author = "Wout Overkamp and Katrin Beilharz and Weme, {Ruud Detert Oude} and Ana Solopova and Harma Karsens and Kovacs, {Akos T.} and Jan Kok and Kuipers, {Oscar P.} and Jan-Willem Veening",

year = "2013",

month = oct,

doi = "10.1128/AEM.02033-13",

language = "English",

volume = "79",

pages = "6481--6490",

journal = "Applied and environmental microbiology",

issn = "0099-2240",

publisher = "AMER SOC MICROBIOLOGY",

number = "20",

}

Overkamp, W, Beilharz, K, Weme, RDO, Solopova, A, Karsens, H, Kovacs, AT, Kok, J , Kuipers, OP & Veening, J-W 2013, 'Benchmarking Various Green Fluorescent Protein Variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for Live Cell Imaging', Applied and environmental microbiology, vol. 79, no. 20, pp. 6481-6490. https://doi.org/10.1128/AEM.02033-13

Benchmarking Various Green Fluorescent Protein Variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for Live Cell Imaging. / Overkamp, Wout; Beilharz, Katrin; Weme, Ruud Detert Oude et al.
In: Applied and environmental microbiology, Vol. 79, No. 20, 10.2013, p. 6481-6490.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Benchmarking Various Green Fluorescent Protein Variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for Live Cell Imaging

AU - Overkamp, Wout

AU - Beilharz, Katrin

AU - Weme, Ruud Detert Oude

AU - Solopova, Ana

AU - Karsens, Harma

AU - Kovacs, Akos T.

AU - Kok, Jan

AU - Kuipers, Oscar P.

AU - Veening, Jan-Willem

PY - 2013/10

Y1 - 2013/10

N2 - 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.

AB - 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.

KW - GENE REGULATORY NETWORKS

KW - QUANTITATIVE-ANALYSIS

KW - OROTATE TRANSPORTER

KW - CODON USAGE

KW - EXPRESSION

KW - COMPETENCE

KW - BACTERIA

KW - PLASMID

KW - GENOME

KW - TRANSFORMATION

U2 - 10.1128/AEM.02033-13

DO - 10.1128/AEM.02033-13

M3 - Article

SN - 0099-2240

VL - 79

SP - 6481

EP - 6490

JO - Applied and environmental microbiology

JF - Applied and environmental microbiology

IS - 20

ER -