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      Permanent link:
      https://www.ias.ac.in/article/fulltext/jbsc/038/04/0713-0717

    • Keywords

       

      Codon usage; Frankia; gentamicin resistance gene; transformation

    • 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 (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.

    • Author Affiliations

       

      Ken-ichi Kucho1 Kentaro Kakoi1 Masatoshi Yamaura1 Mari Iwashita2 Mikiko Abe1 Toshiki Uchiumi1

      1. Graduate School of Science and Engineering, Kagoshima University, Korimoto 1-21-35, Kagoshima 890-0065, Japan
      2. Department of Chemistry and Bioscience, Faculty of Science, Kagoshima University, Korimoto 1-21-35, Kagoshima 890-0065, Japan
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