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

被引：33

作者：

Putz, J

Wientges, J

Sissler, M

Giege, R

Florentz, C

Schwienhorst, A

机构：

[1] INST MOL BIOTECHNOL,D-07745 JENA,GERMANY

[2] CNRS,INST BIOL MOL & CELLULAIRE,UPR 9002,F-67084 STRASBOURG,FRANCE

来源：

NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 09期

关键词：

D O I：

10.1093/nar/25.9.1862

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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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页码：1862 / 1863

页数：2

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