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22 janvier 2021: 1 événement

  • Département Biologie des Génomes

    Vendredi 22 janvier 10:00-11:00 - Masayuki Su'etsugu - Rikkyo University

    In vitro amplification of mega-sized circular DNA and its applications

    Résumé : We have reconstituted an entire replication cycle of the Escherichia coli circular chromosome with 26 proteins that catalyze initiation, fork progression, Okazaki-fragment maturation, and decatenation. Because decatenation provides supercoiled monomers that are competent for the next round of replication initiation, the cycle repeats autonomously and continuously. This Replication-Cycle Reaction (RCR) can propagate large circular DNA up to 1 Mbp. We further developed a multiple-DNA fragment assembly reaction, termed RA (Recombination Assembly). Using a two-step enzymatic reaction, RA-RCR, we have successfully constructed a 27 kb plasmid from 50 fragments. RA-RCR thus provides a powerful cell-free tool to generate large circular DNA without relying on conventional cell-based cloning.


    Contact : Yoshiharu.YAMAICHI

    Lieu : Visio

    En savoir plus : Département Biologie des Génomes