Codon-optimized antibiotic resistance gene improves efficiency of transient transformation in Frankia

Journal of biosciences

Ken-ichi Kucho1¹  Mikiko Abe¹  Mari Iwashita²  Toshiki Uchiumi¹  Kentaro Kakoi¹  Masatoshi Yamaura¹ 

[1] Graduate School of Science and Engineering, Kagoshima University, Korimoto 1-21-35, Kagoshima 890-0065, Japan$$;Department of Chemistry and Bioscience, Faculty of Science, Kagoshima University, Korimoto 1-21-35, Kagoshima 890-0065, Japan$$

关键词: Codon usage;    Frankia;    gentamicin resistance gene;    transformation;   

DOI  :  

来源: Indian Academy of Sciences

【 摘 要 】

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 (fgmR) 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 fgmR 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.

【 授权许可】

Unknown