title: Expression and purification of splicing proteins from mammalian cells.
abstract: Pre-mRNA splicing is a complex process that is carried out by a large ribonucleoprotein enzyme, termed the spliceosome, which comprises up to 200 proteins. Despite this complexity, the role of individual spliceosomal proteins in the splicing reaction has been successfully investigated using cell-free assays. In many cases, the splicing factor of interest must be expressed and purified in order to study its function in vitro. Posttranslational modifications such as phosphorylation, methylation, acetylation, and ubiquitination of splicing factors are important for activity. Thus, their purification from mammalian cells presents numerous advantages. Here, we describe a method for expression and purification of splicing proteins from mammalian cells.
authors.affiliation: Unité de Régulation Epigénétique, Département de Biologie du Développement, Institut Pasteur, Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Allemand
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Hastings
mesh_terms: D000818:Animals; D002474:Cell-Free System; D015870:Gene Expression; D008967:Molecular Biology / Q000379:methods*; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D017381:Spliceosomes / Q000235:genetics*; D054875:Ubiquitination / Q000235:genetics
chemical_list: D011506:Proteins