Rapid selection of aminoacyl-tRNAs based on biotinylation of -NH2 group of charged amino acids | Zendy

Joern Pütz | Zendy; Jens Wientges | Zendy; A. Schwienhorst | Zendy; Marie Sissler | Zendy; R. Giegé | Zendy; C. Florentz | Zendy

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Rapid selection of aminoacyl-tRNAs based on biotinylation of -NH2 group of charged amino acids

Author(s) -

Joern Pütz,

Jens Wientges,

A. Schwienhorst,

Marie Sissler,

R. Giegé,

C. Florentz

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.9.1862

Subject(s) - biotinylation , biology , aminoacylation , streptavidin , transfer rna , biotin , amino acid , biochemistry , amino acyl trna synthetases , selection (genetic algorithm) , rna , gene , artificial intelligence , computer science

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.

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