Cloning, Expression and Characterization of Cyt2aa1 Toxin

A reliable system for expression and purification of the recombinant Cyt2Aa1 toxin has been developed. The recombinant Cyt2Aa1 toxin has been produced, characterized, followed by the construction of the cysteine mutants V186C and L189C by site directed mutagenesis. The hemolytic activity of the V186C mutant exceeds that of wild type Cyt2Aa1 toxin and of the L189C mutant. Calcein release assay experiments have been done to examine the activity of the toxin with different artificial liposomes. It was found that Cyt2Aa1 toxin is very active with DMPC, DMPC+DMPG unilamellar liposomes.