Plasmid transformation of Clostridium perfringens by electroporation methods

Construction of an Escherichia coli-Clostridium perfringens shuttle vector and plasmid transformation of Clostridium perfringens.

A stable shuttle vector which replicates in Escherichia coli and Clostridium perfringens was constructed by ligating a 3.6-kilobase (kb) fragment of plasmid pBR322 with C. perfringens plasmid pHB101 (3.1 kb). The marker for this shuttle plasmid originated from the 1.3-kb chloramphenicol resistance gene of plasmid pHR106. The resulting shuttle vector, designated pAK201, is 8 kb in size and codes...

Transformation of Clostridium thermocellum by electroporation.

In this work, we provide detailed instructions for transformation of Clostridium thermocellum by electroporation. In addition, we describe two schemes for genetic modification: allelic replacement-where the gene of interest is replaced by an antibiotic marker and markerless gene deletion-where the gene of interest is removed and the selective markers are recycled. The markerless gene deletion t...

Transformation of bile acids by Clostridium perfringens.

Thirty-five strains of Clostridium perfringens were examined for their ability to transform bile acids, both in growing cultures and by washed whole cells. All of the strains oxidized the 3 alpha-hydroxy group to an oxo group, and all except three converted the same alpha-hydroxy group into a beta-configuration. The oxidative 3 alpha-dehydrogenation was barely detectable under anaerobic cultura...

Bacteriophages of Clostridium perfringens

Broad-Host-Range Plasmid by Electroporation

Methylotrophic bacteria able to grow on one-carbon compounds, such as methanol and monomethylamine, as sole carbon sources have received considerable attention because of their potential commercial applications, including the production of single-cell protein, amino acids, pyrroloquinoline quinone, and poly-3-hydroxybutyrate. The utility of methylotrophs should be increased by the use of recomb...