Sep 10, 2003 - A custom-built pulse generator was used to ... .home.html) and a number of genes from C. thermocellum have ... E-mail: lee ..... 4); lane 5, pUC18; lane 6, no template; lane 7, pIKm1; lanes 8 to 10, total DNAs of Emr Lmr clones ...
Electrotransformation of Clostridium thermocellum Michael V. Tyurin,1 Sunil G. Desai,1 and Lee R. Lynd1,2* Thayer School of Engineering1 and Department of Biological Sciences,2 Dartmouth College, Hanover, New Hampshire 03755 Received 10 September 2003/Accepted 7 November 2003
Electrotransformation of several strains of Clostridium thermocellum was achieved using plasmid pIKm1 with selection based on resistance to erythromycin and lincomycin. A custom-built pulse generator was used to apply a square 10-ms pulse to an electrotransformation cuvette consisting of a modified centrifuge tube. Transformation was verified by recovery of the shuttle plasmid pIKm1 from presumptive transformants of C. thermocellum with subsequent PCR specific to the mls gene on the plasmid, as well as by retransformation of Escherichia coli. Optimization carried out with strain DSM 1313 increased transformation efficiencies from
