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.
Yanguo Shi and Jinlong Zuo
F. H. Wu et al., "A Novel PCR Site-Directed Mutagenesis to Modify Structure of Erabutoxin B", Advanced Materials Research, Vols. 183-185, pp. 605-610, 2011