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Proc Natl Acad Sci U S A. Feb 28, 1995; 92(5): 1609–1613.
PMCID: PMC42569

Display of peptides and proteins on the surface of bacteriophage lambda.

Abstract

The display of peptides or proteins on the surface of viruses is an important technology for studying peptides or proteins and their interaction with other molecules. Here we describe a display vehicle based on bacteriophage lambda that incorporates a number of features distinct from other currently used display systems. Fusions of peptides or protein domains have been made to the amino terminus of the 11-kDa D protein of the lambda capsid. These fusions assemble onto the viral capsid and appear to be accessible to ligand interactions, based on the ability of a monoclonal antibody to recognize an epitope fused to the D protein on phage heads. To produce large D fusion display libraries and yet avoid the cumbersome task of cloning many fragments into lambda DNA, we have used the Cre-loxP site-specific recombination system in vivo to incorporate plasmids encoding the D fusions into the phage genome. Finally, we show that D fusion proteins can be added in vitro to phage lacking D protein and be assembled onto the viral capsid.

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Selected References

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  • Smith GP. Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science. 1985 Jun 14;228(4705):1315–1317. [PubMed]
  • Parmley SF, Smith GP. Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene. 1988 Dec 20;73(2):305–318. [PubMed]
  • Clackson T, Wells JA. In vitro selection from protein and peptide libraries. Trends Biotechnol. 1994 May;12(5):173–184. [PubMed]
  • Little M, Fuchs P, Breitling F, Dübel S. Bacterial surface presentation of proteins and peptides: an alternative to phage technology? Trends Biotechnol. 1993 Jan;11(1):3–5. [PubMed]
  • Sternberg N, Weisberg R. Packaging of coliphage lambda DNA. II. The role of the gene D protein. J Mol Biol. 1977 Dec 15;117(3):733–759. [PubMed]
  • Imber R, Tsugita A, Wurtz M, Hohn T. Outer surface protein of bacteriophage lambda. J Mol Biol. 1980 May 25;139(3):277–295. [PubMed]
  • Dokland T, Murialdo H. Structural transitions during maturation of bacteriophage lambda capsids. J Mol Biol. 1993 Oct 20;233(4):682–694. [PubMed]
  • Maruyama IN, Maruyama HI, Brenner S. Lambda foo: a lambda phage vector for the expression of foreign proteins. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8273–8277. [PMC free article] [PubMed]
  • Shimada K, Weisberg RA, Gottesman ME. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. [PubMed]
  • Mousset S, Thomas R. Dilysogenic excision: an accessory expression of the termination function? Cold Spring Harb Symp Quant Biol. 1968;33:749–754. [PubMed]
  • O'Neil KT, Hoess RH, Raleigh DP, DeGrado WF. Thermodynamic genetics of the folding of the B1 immunoglobulin-binding domain from streptococcal protein G. Proteins. 1995 Jan;21(1):11–21. [PubMed]
  • Abremski K, Hoess R, Sternberg N. Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination. Cell. 1983 Apr;32(4):1301–1311. [PubMed]
  • Guzman LM, Barondess JJ, Beckwith J. FtsL, an essential cytoplasmic membrane protein involved in cell division in Escherichia coli. J Bacteriol. 1992 Dec;174(23):7716–7728. [PMC free article] [PubMed]
  • Schägger H, von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. [PubMed]
  • Reilly TM, Chiu AT, Timmermans PB. Monoclonal antibodies to angiotensin II. Biochem Biophys Res Commun. 1987 Feb 27;143(1):133–139. [PubMed]
  • Hoess RH, Ziese M, Sternberg N. P1 site-specific recombination: nucleotide sequence of the recombining sites. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3398–3402. [PMC free article] [PubMed]
  • Frick IM, Wikström M, Forsén S, Drakenberg T, Gomi H, Sjöbring U, Björck L. Convergent evolution among immunoglobulin G-binding bacterial proteins. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8532–8536. [PMC free article] [PubMed]
  • Greenwood J, Willis AE, Perham RN. Multiple display of foreign peptides on a filamentous bacteriophage. Peptides from Plasmodium falciparum circumsporozoite protein as antigens. J Mol Biol. 1991 Aug 20;220(4):821–827. [PubMed]
  • Willis AE, Perham RN, Wraith D. Immunological properties of foreign peptides in multiple display on a filamentous bacteriophage. Gene. 1993 Jun 15;128(1):79–83. [PubMed]
  • Hart SL, Knight AM, Harbottle RP, Mistry A, Hunger HD, Cutler DF, Williamson R, Coutelle C. Cell binding and internalization by filamentous phage displaying a cyclic Arg-Gly-Asp-containing peptide. J Biol Chem. 1994 Apr 29;269(17):12468–12474. [PubMed]

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