title: Codon-optimized antibiotic resistance gene improves efficiency of transient transformation in Frankia.
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 (more than 70 percent) 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.
authors.affiliation: Graduate School of Science and Engineering, Kagoshima University, Korimoto 1-21-35, Kagoshima 890-0065, Japan, kkucho@sci.kagoshima-u.ac.jp.
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Kucho
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kakoi
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaura
authors.forename: Mari
authors.initials: M
authors.lastname: Iwashita
authors.forename: Mikiko
authors.initials: M
authors.lastname: Abe
authors.forename: Toshiki
authors.initials: T
authors.lastname: Uchiumi
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001482:Base Composition; D001483:Base Sequence; D003062:Codon / Q000737:chemistry*; D003470:Culture Media; D004352:Drug Resistance, Microbial / Q000235:genetics; D040161:Frankia / Q000187:drug effects / Q000235:genetics*; D005818:Genetic Engineering; D016680:Genome, Bacterial; D005839:Gentamicins / Q000494:pharmacology; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation / Q000502:physiology; D010944:Plants / Q000382:microbiology; D010957:Plasmids / Q000737:chemistry; D053204:Root Nodules, Plant / Q000382:microbiology / Q000502:physiology; D013559:Symbiosis; D014169:Transformation, Bacterial; D019076:Transgenes
chemical_list: D000900:Anti-Bacterial Agents; D003062:Codon; D003470:Culture Media; D005839:Gentamicins