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  • Title: Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM 792.
    Author: Nakotte S, Schaffer S, Böhringer M, Dürre P.
    Journal: Appl Microbiol Biotechnol; 1998 Nov; 50(5):564-7. PubMed ID: 9866174.
    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.
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