Molecular Biotechnology

, Volume 46, Issue 1, pp 58–62

Characterisation of a DNA Polymerase Highly Mutated Along the Template Binding Interface


DOI: 10.1007/s12033-010-9275-4

Cite this article as:
Vichier-Guerre, S. & Jestin, JL. Mol Biotechnol (2010) 46: 58. doi:10.1007/s12033-010-9275-4


Phage display establishes a link between a polypeptide and its corresponding gene. It has been much used for the isolation of proteins binding to chosen molecular targets. A second link was designed more recently between a phage-displayed enzyme and its reaction product. Affinity chromatography for the product then allows the isolation of catalytically active enzymes and of their genes. Using this strategy, a polymerase with 15 mutations was selected by directed evolution of Thermus aquaticus DNA polymerase I. The kinetic characterisation reported here highlights the variant’s broad template specificity and classifies this enzyme as a thermostable DNA-dependent and RNA-dependent DNA-polymerase.


Reverse transcriptase Enzyme engineering In vitro selection Phage display Directed evolution 

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Département de Biologie Structurale et ChimieInstitut PasteurParis 15France

Personalised recommendations