SERF: in vitro election of random RNA fragments to identify protein binding sites within large RNAs.
In vitro selection experiments have various goals depending on the composition of the initial pool and the selection method applied. We developed an in vitro selection variant (SERF, selection of random RNA fragments) that is useful for the identification of short RNA fragments originating from large RNAs that bind specifically to a protein. A pool of randomly fragmented RNA is constructed from a large RNA, which is the natural binding partner for a protein. Such a pool contains all the potential binding sites and is therefore used as starting material for affinity selection with the purified protein to find its natural target. Here we provide a detailed experimental protocol of the method. SERF has been developed for ribosomal systems and is a general approach providing a basis for functional and structural characterization of RNA-protein interactions in large ribonucleoprotein particles.
Stelzl
U
U
Max-Planck-Institut für Molekulare Genetik, AG Ribosomen, Ihnestrasse 73, D-14195 Berlin, Germany.
Nierhaus
K H
KH
D001419:Bacteria / Q000378:metabolism; D001665:Binding Sites; D003101:Collodion / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure*; D005821:Genetic Techniques; D008957:Models, Genetic; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry* / Q000648:ultrastructure; D012335:RNA, Ribosomal / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors
D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D012313:RNA; D003101:Collodion; D004247:DNA