Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM 792 |
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Authors: | S Nakotte S Schaffer M B?hringer P Dürre |
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Affiliation: | Angewandte Mikrobiologie und Mykologie, Universit?t Ulm, Germany. |
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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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