An efficient electroporation procedure for Vibrio vulnificus was designed using the new cloning vector pVv3 (3,107 bp). Transformation efficiencies up to 2 × 10(6) transformants per μg DNA were achieved. The vector stably replicated in both V. vulnificus and Escherichia coli and was also successfully introduced into Vibrio parahaemolyticus and Vibrio cholerae. To demonstrate the suitability of the vector for molecular cloning, the green fluorescent protein (GFP) gene and the vvhBA hemolysin operon were inserted into the vector and functionally expressed in Vibrio and E. coli.
We report here the 79,263-bp plasmid pVv01 isolated from Vibrio vulnificus. pVv01 is closely related to the Vibrio plasmid p0908 and shows some similarities to phage P1. Unlike p0908, pVv01 represents an intact prophage inducible by mitomycin C. PVv01 phage particles revealed a myoviridal morphology and lytic activity.
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