title: Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM 792.
abstract: Procedures have been developed allowing recombinant DNA work with Clostridium acetobutylicum DSM 792. Electroporation was used to introduce plasmid DNA into exponentially growing clostridial cells and 6 x 10(2) transformants/microgram DNA could be obtained at a time constant of 5.5 ms, 1.8 kV, 50 microF, and 600 omega. The method also allowed the taxonomic group IV strain NI-4082 to be transformed (10(1) transformants/microgram DNA). Plasmid preparation from recombinant clostridia was optimal when a modification of the alkaline lysis method was employed. It was also important to use cells from the mid-logarithmic growth phase. Recombinant strains could be easily preserved as spore suspensions; under all conditions tested plasmids were maintained.
authors.affiliation: Angewandte Mikrobiologie und Mykologie, Universität Ulm, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Nakotte
authors.forename: S
authors.initials: S
authors.lastname: Schaffer
authors.forename: M
authors.initials: M
authors.lastname: Böhringer
authors.forename: P
authors.initials: P
authors.lastname: Dürre
mesh_terms: D000693:Anaerobiosis; D003013:Clostridium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D018274:Electroporation / Q000379:methods*; D005818:Genetic Engineering; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification; D013171:Spores, Bacterial; D014169:Transformation, Bacterial