A Novel PCR Site-Directed Mutagenesis to Modify Structure of Erabutoxin B

Objective: A convenient PCR was used for site-directed mutagenesis to modify structure of proteins or peptides, For function-structure studies of proteins or peptides. Method: the one-step PCR site-directed mutagenesis strategy can introduce mutation of gene through 5/-end of primer, which was applied on peptide Erabutoxin B (EB) to produce three mutants of EB, the activity of mutants was detected by LD50 value of mice. Result: S8Y, R33D, K47R three mutants of EB were obtained by one-step PCR, LD50 of mutants indicated that the activity of mutants decreased in different degree, the activity of R33D was nearly deprived. Conclusion: one-step PCR site-directed mutagenesis was convenien and efficient, it can be applied on restructuring the primary structure of proteins or peptides.