Identification of mRNA-binding proteins during development: characterization of Bufo arenarum cellular nucleic acid binding protein.

摘要: Ultraviolet irradiation was used to covalently cross-link poly(A)+RNA and associated proteins in eggs embryos of the toad Bufo arenarum. Four major with apparent sizes 60, 57, 45 30–24 kDa were identified. It observed that same mRNA-binding isolated from gastrula neural stages development. The 30 polypeptide, p30, appeared as main ultraviolet (UV) cross-linked protein developmental analyzed. By means polyclonal antibodies, it determined this polypeptide has a cytoplasmic localization detected liver, embryos. presence p30 also analyzed by western blot during oogenesis present all but could not be I–II oogenesis. At stage, relative amount began decrease, reaching its lowest levels after 26–30 (tail-bud arenarum). On basis purification, immunoprecipitation assays identified arenarum cellular nucleic acid binding protein.