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Journal of microbiology and biotechnology v.20 no.6, 2010년, pp.1022 - 1026   SCIE 피인용횟수: 4
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Improvement of Transformation Efficiency Through In Vitro Methylation and SacII Site Mutation of Plasmid Vector in Bifidobacterium longum MG1

Kim, Jin-Yong    (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University   ); Wang, Yan    (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University   ); Park, Myeong-Soo    (Department of Hotel Culinary Arts, Anyang Technical College   ); Ji, Geun-Eog    (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University  );
  • 초록

    The different cleavage patterns of pYBamy59 plasmid isolated from E. coli $DH5{\alpha}$ and B. longum MG1 by the cell extract of B. longum MG1 suggested that the main reason for its low transformation efficiency was related to the restriction modification (R-M) system. To confirm the correlation between the R-M system and transformation efficiency, in vitro methylation and site-directed mutagenesis were performed in pYBamy59. Sequence analysis of pYBamy59 fragments digested by the cell extract of B. longum MG1 revealed that all fragments were generated by restriction of the sequence recognized by SacII endonuclease. When pYBamy59 from E. coli was methylated in vitro by CpG or GpC methyltransferase, it was protected from SacII digestion. Site-directed mutagenesis, which removed SacII sites from pYBamy59, or in vitro methylation of pYBamy59 showed 8- to 15-fold increases in the transformation efficiency over intact pYBamy59. Modification of the SacII-related R-M system in B. longum MG1 and in vitro methylation in pYBamy 59 can improve the transformation efficiency in this strain. The results showed that the R-M system is a factor to limit introduction of exogenous DNA, and in vitro modification is a convenient method to overcome the barrier of the R-M system for transformation.


  • 주제어

    Bifidobacterium .   transformation efficiency .   methylation .   mutagenesis .   SacII.  

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  • 이 논문을 인용한 문헌 (4)

    1. Park, Myeong-Soo ; Ji, Geun-Eog 2010. "Research Trends in Bifidobacterium" KSBB Journal, 25(4): 319~329     
    2. 2012. "" Journal of microbiology and biotechnology, 22(12): 1714~1723     
    3. 2012. "" Food science and biotechnology, 21(3): 731~738     
    4. 2015. "" Journal of microbiology and biotechnology, 25(4): 469~478     

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