Fluorescent T7 display phages obtained by translational frameshift

E.J. Slootweg, H.J.H.G. Keller, M.A. Hink, J.W. Borst, J. Bakker, A. Schots

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)

Abstract

Lytic phages form a powerful platform for the display of large cDNA libraries and offer the possibility to screen for interactions with almost any substrate. To visualize these interactions directly by fluorescence microscopy, we constructed fluorescent T7 phages by exploiting the flexibility of phages to incorporate modified versions of its capsid protein. By applying translational frameshift sequences, helper plasmids were constructed that expressed a fixed ratio of both wild-type capsid protein (gp10) and capsid protein fused to enhanced yellow fluorescent protein (EYFP). The frameshift sequences were inserted between the 3' end of the capsid gene and the sequence encoding EYFP. Fluorescent fusion proteins are only formed when the ribosome makes a ¿1 shift in reading frame during translation. Using standard fluorescence microscopy, we could sensitively monitor the enrichment of specific binders in a cDNA library displayed on fluorescent T7 phages. The perspectives of fluorescent display phages in the fast emerging field of single molecule detection and sorting technologies are discussed
Original languageEnglish
Article numbere137
Number of pages11
JournalNucleic acids research
Volume34
Issue number20
DOIs
Publication statusPublished - 2006

Keywords

  • dye-labeled bacteriophages
  • correlation spectroscopy
  • escherichia-coli
  • viral transport
  • porous-media
  • protein
  • peptide
  • expression
  • antibodies
  • molecules

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