title: Rapid selection of aminoacyl-tRNAs based on biotinylation of alpha-NH2 group of charged amino acids.
abstract: A rapid selection procedure to separate low amounts of aminoacylated tRNAs from large pools of inactive variants is described. The procedure involves a three-step protocol. After initial aminoacylation of a tRNA pool, N-hydroxysuccinimide ester chemistry is applied to biotinylate the alpha-NH2 group of the amino acid bound to the 3'-end of a tRNA. The biotin tag is used to capture the derivatized tRNAs on streptavidin-conjugated magnetic beads. Variants bound to the solid phase can be amplified by RT-PCR and transcription, providing tRNAs for subsequent selection rounds.
authors.affiliation: Institüt für Molekulare Biotechnologie, Beutenbergstrasse 11, D-07745, Jena, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Pütz
authors.forename: J
authors.initials: J
authors.lastname: Wientges
authors.forename: M
authors.initials: M
authors.lastname: Sissler
authors.forename: R
authors.initials: R
authors.lastname: Giegé
authors.forename: C
authors.initials: C
authors.lastname: Florentz
authors.forename: A
authors.initials: A
authors.lastname: Schwienhorst
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D000596:Amino Acids; D012346:RNA, Transfer, Amino Acyl; D001710:Biotin