title: Cloning of a cDNA encoding an RNA binding protein by screening expression libraries using a northwestern strategy.
abstract: Successful cloning with cDNA expression libraries involves interaction of ligand molecules (probes) with expressed fusion proteins of interest. So far those ligands leading to successful results fall into three classes: (i) antibodies, (ii) protein ligands that interact with the protein of interest, and (iii) DNA sequences recognized by transcription factors. We have previously identified a 50-kDa protein (called DSEF-1) which interacts with a functionally important 14-base G-rich RNA sequence located downstream of the simian virus 40 late polyadenylation signal. By using small RNAs containing the DSEF-1 binding site as probes, a cDNA clone was isolated whose gene product interacted in a sequence-specific fashion with the DSEF-1 binding site. This RNA binding protein contains three potential RNA recognition motifs. We present here a procedure to obtain cDNA clones of RNA binding proteins using recognition site probes.
authors.affiliation: Department of Microbiology & Molecular Genetics, UMD-New Jersey Medical School, Newark 07103.
authors.forename: Z
authors.initials: Z
authors.lastname: Qian
authors.forename: J
authors.initials: J
authors.lastname: Wilusz
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003101:Collodion; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D015347:RNA Probes; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D008567:Membranes, Artificial; D015347:RNA Probes; D016601:RNA-Binding Proteins; D003101:Collodion