An efficient transposon mutagenesis system using conjugative transposons Tn916 and Tn925::Tn917 was established for Clostridium acetobutylicum P262, an industrial strain which has proved difficult to manipulate genetically. Transposon insertions occurred at several different locations to produce a variety of mutants. An oligosporogenous mutant deficient in acetone and butanol production, and two sporulation-deficient and metronidazole resistant mutants were characterized with respect to differentiation and solvent production. Tn925::Tn917 inserted near a string of adenosine residues and transposon insertion was often multiple.
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