PMID: 11316373Apr 24, 2001Paper

Improving the display of proteins on filamentous phage

Research in Microbiology
J L JestinG Winter

Abstract

In phage display technology, polypeptides are displayed on the surface of filamentous bacteriophage by genetic fusion to a coat protein. However, the fraction of phage particles bearing the fusion protein can be low. Here we found that we could improve the display of a protein (Stoffel fragment of Taq polymerase fused to the p3 protein of the phage) by mutation of the signal sequence and use of helper phage with a protease-cleavable coat protein. Over multiple rounds of infection, proteolysis and binding to an anti-Taq antibody, we were able to select strongly for display of the fusion protein (> 50-fold), and for mutations in the translation initiation region and in the signal sequence of the fusion. This suggests a general means of improving the display of proteins on phage.

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Citations

Dec 17, 2009·Molecular Biotechnology·Tuomas HuovinenUrpo Lamminmäki
Apr 22, 2010·Molecular Biotechnology·Sophie Vichier-Guerre, Jean-Luc Jestin
Jun 27, 2002·Biochemical and Biophysical Research Communications·Hideki WatanabeIzumi Kumagai
Sep 2, 2003·Trends in Biotechnology·Ana Fernandez-GacioJacques Fastrez
Apr 20, 2004·Assay and Drug Development Technologies·Simon Delagrave, Dennis J Murphy
Nov 8, 2006·Nucleic Acids Research·L ChasteenA R M Bradbury
Jul 29, 2015·Proceedings of the National Academy of Sciences of the United States of America·Changming YuMark J Federspiel
Sep 24, 2004·Journal of Biotechnology·Jean-Luc Jestin, Pierre Alexandre Kaminski
Jul 21, 2004·Journal of Immunological Methods·Andrew R M Bradbury, James D Marks
May 17, 2005·Biochimica Et Biophysica Acta·Eeva-Christine BrockmannPetri Saviranta
Dec 1, 2001·Expert Opinion on Biological Therapy·P J Hudson, C Souriau
Apr 29, 2005·Biomolecular Engineering·Rebecca C HolmbergFloyd E Romesberg
Feb 19, 2011·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yuliya Georgieva, Zoltán Konthur
Sep 7, 2006·The Journal of Gene Medicine·Daniel A WaterkampJürgen A Kleinschmidt
Sep 6, 2007·Journal of Cellular Biochemistry·Christian C KazaneckiDavid T Denhardt
Mar 3, 2010·Cellular and Molecular Life Sciences : CMLS·Tomaz Bratkovic
Jul 31, 2009·Chemical Communications : Chem Comm·David Loakes, Philipp Holliger
Sep 3, 2020·International Journal of Molecular Sciences·Irina A IshinaAlexey A Belogurov

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