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Site-Directed Mutagenesis on Plasmid Using Polymerase Chain Reaction

  • Abdul Aziz Ahmad
  • Hamzah Mohd SallehEmail author
  • Ibrahim Ali Noorbatcha
Chapter

Abstract

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.

Keywords

Site directed mutagenesis Endoglucanase Fusarium oxysporum 

Notes

Acknowledgement

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.

References

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Copyright information

© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  • Abdul Aziz Ahmad
    • 1
  • Hamzah Mohd Salleh
    • 1
    • 2
    Email author
  • Ibrahim Ali Noorbatcha
    • 1
  1. 1.BioProcess and Molecular Engineering Research Unit, Kulliyyah of EngineeringInternational Islamic University MalaysiaKuala LumpurMalaysia
  2. 2.International Institute for Halal Research and Training, International Islamic University MalaysiaKuala LumpurMalaysia

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