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 Resjournal_title
Nucleic acids researchauthors
Cox JC,Hayhurst A,Hesselberth J,Bayer TS,Georgiou G,Ellington ADdoi
10.1093/nar/gnf107keywords:
subject
Has Abstractpub_date
2002-10-15 00:00:00pages
e108issue
20eissn
0305-1048issn
1362-4962journal_volume
30pub_type
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