On-bead expression of recombinant proteins in an agarose gel matrix coated on a glass slide

Lab Chip. 2012 May 7;12(9):1605-10. doi: 10.1039/c2lc21239k. Epub 2012 Mar 12.

Abstract

A system for expression and in situ display of recombinant proteins on a microbead surface is described. Biotinylated PCR products were immobilized on microbead surfaces, which were then embedded in a gel matrix and supplied with translation machinery and substrates. Upon the incubation of the gel matrix, target proteins encoded on the bead-immobilized DNA were expressed and captured on the same bead, thus allowing bead-mediated linkage of DNA and encoded proteins. The new method combines the simplicity and convenience of solid-phase separation of genetic information with the benefits of cell-free protein synthesis, such as instant translation of genetic information, unrestricted substrate accessibility and flexible assay configuration design.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin
  • Cell-Free System / metabolism*
  • DNA / genetics*
  • Glass
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Microscopy, Confocal
  • Microspheres*
  • Polymerase Chain Reaction
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sepharose / chemistry*

Substances

  • Recombinant Proteins
  • Green Fluorescent Proteins
  • Biotin
  • DNA
  • Sepharose
  • Luciferases