title: On-bead expression of recombinant proteins in an agarose gel matrix coated on a glass slide.
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.
authors.affiliation: Interdisciplinary Program for Nano-Technology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Kyung-Ho
authors.initials: KH
authors.lastname: Lee
authors.forename: Ka-Young
authors.initials: KY
authors.lastname: Lee
authors.forename: Ju-Young
authors.initials: JY
authors.lastname: Byun
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
mesh_terms: D001710:Biotin; D002474:Cell-Free System / Q000378:metabolism*; D004247:DNA / Q000235:genetics*; D005898:Glass; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D018613:Microscopy, Confocal; D008863:Microspheres; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012685:Sepharose / Q000737:chemistry*
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D001710:Biotin; D004247:DNA; D012685:Sepharose; D008156:Luciferases