Site-Directed Mutagenesis on Plasmid Using Polymerase Chain Reaction
Site-directed mutagenesis (SDM) is a very useful technique to study changes in protein function that may occur as a result of the DNA manipulation. A detail procedure to employ SDM on whole plasmid using polymerase chain reaction (PCR) with successful application of the method on a pET28a plasmid harboring an endoglucanase I gene from Fusarium oxysporum is described here.
KeywordsSite directed mutagenesis Endoglucanase Fusarium oxysporum
The authors would like to acknowledge the Ministry of Higher Education, (MoHE) Malaysia for awarding fund via Fundamental Research Grant Scheme: Grant No: FRGS-13-070-0311 and Professor S.G. Withers (UBC, Vancouver) for providing the original wild-type plasmid construct.