Automated selection of aptamers against protein targets translated in vitro: from gene to aptamer.

Abstract:

:Reagents for proteome research must of necessity be generated by high throughput methods. Aptamers are potentially useful as reagents to identify and quantitate individual proteins, yet are currently produced for the most part by manual selection procedures. We have developed automated selection methods, but must still individually purify protein targets. Therefore, we have attempted to select aptamers against protein targets generated by in vitro transcription and translation of individual genes. In order to specifically immobilize the protein targets for selection, they are also biotinylated in vitro. As a proof of this method, we have selected aptamers against translated human U1A, a component of the nuclear spliceosome. Selected sequences demonstrated exquisite mimicry of natural binding sequences and structures. These results not only reveal a potential path to the high throughput generation of aptamers, but also yield insights into the incredible specificity of the U1A protein for its natural RNA ligands.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Cox JC,Hayhurst A,Hesselberth J,Bayer TS,Georgiou G,Ellington AD

doi

10.1093/nar/gnf107

keywords:

subject

Has Abstract

pub_date

2002-10-15 00:00:00

pages

e108

issue

20

eissn

0305-1048

issn

1362-4962

journal_volume

30

pub_type

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