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
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.
引用
收藏
页码:1862 / 1863
页数:2
相关论文
共 12 条
[1]   ACYLATION OF TRANSFER RIBONUCLEIC-ACID WITH N-HYDROXYSUCCINIMIDE ESTER OF PHENOXYACETIC ACID [J].
FRIEDMAN, S .
BIOCHEMISTRY, 1972, 11 (18) :3435-&
[2]   DIVERSITY OF OLIGONUCLEOTIDE FUNCTIONS [J].
GOLD, L ;
POLISKY, B ;
UHLENBECK, O ;
YARUS, M .
ANNUAL REVIEW OF BIOCHEMISTRY, 1995, 64 :763-797
[3]   AMINOACYL-RNA SYNTHESIS CATALYZED BY AN RNA [J].
ILLANGASEKARE, M ;
SANCHEZ, G ;
NICKLES, T ;
YARUS, M .
SCIENCE, 1995, 267 (5198) :643-647
[4]   TRANSFER RNA-MEDIATED LABELING OF PROTEINS WITH BIOTIN A NONRADIOACTIVE METHOD FOR THE DETECTION OF CELL-FREE TRANSLATION PRODUCTS [J].
KURZCHALIA, TV ;
WIEDMANN, M ;
BRETER, H ;
ZIMMERMANN, W ;
BAUSCHKE, E ;
RAPOPORT, TA .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 172 (03) :663-668
[5]   Ribozyme-catalysed amino-acid transfer reactions [J].
Lohse, PA ;
Szostak, JW .
NATURE, 1996, 381 (6581) :442-444
[6]   SELECTION FOR ACTIVE ESCHERICHIA-COLI TRANSFER RNA(PHE) VARIANTS FROM A RANDOMIZED LIBRARY USING 2 PROTEINS [J].
PETERSON, ET ;
BLANK, J ;
SPRINZL, M ;
UHLENBECK, OC .
EMBO JOURNAL, 1993, 12 (07) :2959-2967
[7]   IDENTITY ELEMENTS FOR SPECIFIC AMINOACYLATION OF YEAST TRANSFER RNAASP BY COGNATE ASPARTYL-TRANSFER RNA-SYNTHETASE [J].
PUTZ, J ;
PUGLISI, JD ;
FLORENTZ, C ;
GIEGE, R .
SCIENCE, 1991, 252 (5013) :1696-1699
[8]   PURIFICATION OF AMINOACYL-TRANSFER-RNA BY AFFINITY-CHROMATOGRAPHY ON IMMOBILIZED THERMUS-THERMOPHILUS EF-TU-CENTER-DOT-GTP [J].
RIBEIRO, S ;
NOCK, S ;
SPRINZL, M .
ANALYTICAL BIOCHEMISTRY, 1995, 228 (02) :330-335
[9]  
Riordan J F, 1972, Methods Enzymol, V25, P449, DOI 10.1016/S0076-6879(72)25040-6
[10]  
Sampson JR, 1996, METHOD ENZYMOL, V267, P384