RESUMEN
The PRP4 (RNA4) gene product is involved in nuclear mRNA processing in yeast cells; we have previously cloned the gene by complementation of a temperature-sensitive mutation. Sequence and transcript analyses of the cloned gene predicted the gene product to be a 52-kilodalton protein, which was confirmed with antibodies raised against the PRP4 gene product. These antibodies inhibited precursor mRNA splicing in vitro, demonstrating a direct role of PRP4 in splicing. Immunoprecipitations with the antibodies indicated that the PRP4 protein is associated with the U4/U6 small nuclear ribonucleoprotein particle.
Asunto(s)
Genes Fúngicos , Ribonucleoproteínas/genética , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , Anticuerpos Antifúngicos , Secuencia de Bases , ADN de Hongos/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/inmunología , Datos de Secuencia Molecular , Empalme del ARN , Ribonucleoproteínas/inmunología , Ribonucleoproteínas Nucleares Pequeñas , Transcripción GenéticaRESUMEN
Use of the green fluorescent protein (Gfp) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition. To expand the use of Gfp as a reporter protein, new variants have been constructed by the addition of short peptide sequences to the C-terminal end of intact Gfp. This rendered the Gfp susceptible to the action of indigenous housekeeping proteases, resulting in protein variants with half-lives ranging from 40 min to a few hours when synthesized in Escherichia coli and Pseudomonas putida. The new Gfp variants should be useful for in situ studies of temporal gene expression.