Abstract
This chapter discusses reliable and reproducible methods for the transformation of Clostridium perfringens cells with recombinant plasmids and constructs several well characterized shuttle vectors. Methods for transposon mutagenesis and homologous recombination also have been elucidated and used for the functional analysis of C. perfringens genes. The first method developed for the transformation of C. perfringens cells involved the transformation of L-phase variants generated by growth in the presence of penicillin. Transposon mutagenesis has been widely used for the genetic analysis of both chromosomal and plasmid-determined genes. The only transposons reported from C. perfringens are the chloramphenicol resistance transposons Tn4451 and Tn4452. The tools available for genetic analysis in C. perfringens have been developed to the stage that they are equivalent to many other Gram-positive bacteria. Clostridium perfringens has the well developed genetic system of the pathogenic clostridia and as such are considered as the paradigm bacterium for the functional genetic analysis of the pathogenesis of toxaemic clostridial diseases.
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