Abstract
Frankia is a unique actinobacterium having abilities to fix atmospheric dinitrogen and to establish endosymbiosis with trees, but molecular bases underlying these interesting characteristics are poorly understood because of a lack of stable transformation system. Extremely high GC content of Frankia genome (>70%) can be a hindrance to successful transformation. We generated a synthetic gentamicin resistance gene whose codon usage is optimized to Frankia (fgm R) and evaluated its usefulness as a selection marker using a transient transformation system. Success rate of transient transformation and cell growth in selective culture were significantly increased by use of fgm R instead of a native gentamicin resistance gene, suggesting that codon optimization improved translation efficiency of the marker gene and increased antibiotic resistance. Our result shows that similarity in codon usage pattern is an important factor to be taken into account when exogenous transgenes are expressed in Frankia cells.
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Acknowledgements
We thank Dr Louis S Tisa (University of New Hampshire) for generously providing the Frankia sp. strain CcI3. This work was supported by grants from Nissan Science Foundation, The Asahi Glass Foundation and Kagoshima Kagaku Kenkyuusyo, and MEXT KAKENHI 21770053.
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[Kucho K, Kakoi K, Yamaura M, Iwashita M, Abe M and Uchiumi T 2013 Codon-optimized antibiotic resistance gene improves efficiency of transient transformation in Frankia. J. Biosci. 38 1–5] DOI 10.1007/s12038-013-9361-4
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Kucho, Ki., Kakoi, K., Yamaura, M. et al. Codon-optimized antibiotic resistance gene improves efficiency of transient transformation in Frankia . J Biosci 38, 713–717 (2013). https://doi.org/10.1007/s12038-013-9361-4
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DOI: https://doi.org/10.1007/s12038-013-9361-4