Electroporation of, plasmid isolation from and plasmid conservation in Clostridium acetobutylicum DSM 792.
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.
Nakotte
S
S
Angewandte Mikrobiologie und Mykologie, Universität Ulm, Germany.
Schaffer
S
S
Böhringer
M
M
Dürre
P
P
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