Purification or RNA and RNA—protein complexes by an R17 coat protein affinity method

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Purification of RNA and RNA-protein complexes by an R17 coat protein affinity method.

We describe an affinity chromatography method to isolate specific RNAs and RNA-protein complexes formed in vivo or in vitro. It exploits the highly selective binding of the coat protein of bacteriophage R17 to a short hairpin in its genomic RNA. RNA containing that hairpin binds to coat protein that has been covalently bound to a solid support. Bound RNA-protein complexes can be eluted with exc...

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Affinity purification of protein complexes.

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RNA affinity tags for purification of RNAs and ribonucleoprotein complexes.

Intrinsic affinity tags are useful tools for the study of macromolecular targets. Although polypeptide affinity tags are routinely used in purification and detection of protein complexes, there has been a relative lack of powerful RNA affinity tags that can be embedded within RNA sequences. Here, the preparation and use of two RNA affinity tags against Sephadex or streptavidin are described. Th...

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The tandem affinity purification method: an efficient system for protein complex purification and protein interaction identification.

Isolation and identification of protein partners in multi-protein complexes are important in gaining further insights into the cellular roles of proteins and determining the possible mechanisms by which proteins have an effect in the molecular environment. The tandem affinity purification (TAP) method was originally developed in yeast for the purification of protein complexes and identification...

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Protein Purification by Affinity Chromatography

The preparation of a number of agarose and polyacrylamide bead derivatives useful in the purification of proteins by afhnity chromatography is described. These techniques permit (a) the attachment of ligands to the gel through extended hydrocarbon chains which place the ligand at varying distances from the gel matrix backbone; (b) the covalent attachment of ligands to agarose or polyacrylamide ...

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 1991

ISSN: 0305-1048,1362-4962

DOI: 10.1093/nar/19.8.1980-a