Efficient phage display of polypeptides fused to the carboxy-terminus of the M13 gene-3 minor coat protein.
We report that, contrary to common belief, polypeptides fused to the carboxy-terminus of the M13 gene-3 minor coat protein are functionally displayed on the phage surface. In a phagemid display system, carboxy-terminal fusion through optimized linker sequences resulted in display levels comparable to those achieved with conventional amino-terminal fusions. These findings are of considerable importance to phage display technology because they enable investigations not suited to amino-terminal display, including the study of protein-protein interactions requiring free carboxy-termini, functional cDNA cloning efforts, and the display of intracellular proteins.
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Department of Protein Engineering, Genetech, Inc, South San Francisco, CA 94080, USA.
Sidhu
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D000595:Amino Acid Sequence; D019677:Artificial Gene Fusion; D017104:Bacteriophage M13 / Q000235:genetics; D001665:Binding Sites; D036022:Capsid Proteins; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D016228:Endothelial Growth Factors / Q000378:metabolism; D008222:Lymphokines / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1; D042442:Vascular Endothelial Growth Factors; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D016228:Endothelial Growth Factors; D008222:Lymphokines; D019151:Peptide Library; D010455:Peptides; D011518:Proto-Oncogene Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014760:Viral Fusion Proteins; D020794:Receptor Protein-Tyrosine Kinases; D040281:Vascular Endothelial Growth Factor Receptor-1