Improving the display of proteins on filamentous phage.
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.
Jestin
J L
JL
Centre for Protein Engineering and Laboratory of Molecular Biology, Medical Research Council Centre, Cambridge, UK. jjestin@pasteur.fr
Volioti
G
G
Winter
G
G
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D019151:Peptide Library; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins; D012641:Selection, Genetic; D019914:Taq Polymerase / Q000032:analysis / Q000737:chemistry / Q000235:genetics
D019151:Peptide Library; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D019914:Taq Polymerase