SELECTION OF HIGH-AFFINITY RNA LIGANDS TO THE BACTERIOPHAGE-R17 COAT PROTEIN

被引:121
作者
SCHNEIDER, D
TUERK, C
GOLD, L
机构
[1] University of Colorado at Boulder Campus Box 347
关键词
SELEX; R17-COAT PROTEIN; RNA HAIRPIN; RNA LIGAND; DISSOCIATION CONSTANT;
D O I
10.1016/0022-2836(92)90870-P
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
RNA ligands with high affinity for the bacteriophage R17 coat protein were isolated from a pool of random RNA molecules using SELEX. Of the 38 ligands isolated, 36 were found to contain a hairpin very similar to the naturally occurring coat protein binding site in the R17 genome. The common features of these 36 sequences provide a consensus binding site and predict components of a hairpin that promote favorable interaction with the coat protein. These include a tetraloop of primary sequence AUCA and a variable-length stem with a bulged adenosine residue at a specific stem position. The predicted consensus agrees well with the highest-affinity RNA binding site of the R17 coat protein, identified through classical but laborious techniques. These results demonstrate the value of SELEX as a tool for isolating high affinity RNA ligands to a specific target protein, and the further value of those ligands to point the researcher toward natural sequences for that target protein. © 1992.
引用
收藏
页码:862 / 869
页数:8
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