Construction of a filamentous phage display peptide library

Methods Mol Biol. 2014:1088:19-33. doi: 10.1007/978-1-62703-673-3_2.

Abstract

The concept of phage display is based on insertion of random oligonucleotides at an appropriate location within a structural gene of a bacteriophage. The resulting phage will constitute a library of random peptides displayed on the surface of the bacteriophages, with the encoding genotype packaged within each phage particle. Using a phagemid/helper phage system, the random peptides are interspersed between wild-type coat proteins. Libraries of phage-expressed peptides may be used to search for novel peptide ligands to target proteins. The success of finding a peptide with a desired property in a given library is highly dependent on the diversity and quality of the library. The protocols in this chapter describe the construction of a high-diversity library of phagemid vector encoding fusions of the phage coat protein pVIII with random peptides, from which a phage library displaying random peptides can be prepared.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Genetic Vectors / metabolism
  • Inovirus / metabolism*
  • Molecular Sequence Data
  • Oligonucleotides / metabolism
  • Peptide Library*
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Transformation, Genetic

Substances

  • Oligonucleotides
  • Peptide Library