Conjugative transfer of the Escherichia coli-Clostridium perfringens shuttle vector pJIR1457 to Clostridium botulinum type A strains.
An RP4-oriT shuttle vector pJIR1457 originally developed for Clostridium perfringens was successfully transferred by conjugation from Escherichia coli to Clostridium botulinum type A strains and to a nontoxigenic C. botulinum type A-transposon Tn916 mutant strain lacking the entire toxin gene cluster. The light chain (LC) of botulinum toxin was highly expressed in the toxin deletion mutant strain from a pJIR1457 construct containing the recombinant botulinal gene for LC. This shuttle vector system will be valuable for genetic analysis of C. botulinum and will enable genetic manipulation and recombinant expression studies of botulinum neurotoxins as pharmaceutical agents.
Bradshaw
M
M
Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, Wisconsin, 53706, USA.
Goodnough
M C
MC
Johnson
E A
EA
D001905:Botulinum Toxins / Q000096:biosynthesis / Q000235:genetics; D003014:Clostridium botulinum / Q000235:genetics*; D003016:Clostridium perfringens / Q000235:genetics*; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D004251:DNA Transposable Elements; D011993:Recombinant Fusion Proteins; D001905:Botulinum Toxins