title: Preparation of a Saccharomyces cerevisiae cell-free extract for in vitro translation.
abstract: Eukaryotic cell-free in vitro translation systems have been in use since the 1970s. These systems can faithfully synthesize polypeptides when programmed with mRNA, enabling the production of polypeptides for analysis as well as permitting analyses of the cis- and trans-acting factors that regulate translation. Here we describe the preparation and use of cell-free translation systems from the yeast Saccharomyces cerevisiae.
authors.affiliation: Department of Biology, Texas A&M University, College Station, TX, USA.
authors.forename: Cheng
authors.initials: C
authors.lastname: Wu
authors.affiliation: Department of Biology, Texas A&M University, College Station, TX, USA. Electronic address: msachs@mail.bio.tamu.edu.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Sachs
mesh_terms: D002021:Buffers; D002457:Cell Extracts / Q000302:isolation & purification*; D002458:Cell Fractionation; D017930:Genes, Reporter; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D011480:Protease Inhibitors / Q000737:chemistry; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000166:cytology; D012996:Solutions
chemical_list: D002021:Buffers; D002457:Cell Extracts; D011480:Protease Inhibitors; D012996:Solutions; D049410:Luciferases, Renilla
keywords: Amino acids; Cell-free translation; Chromatography; Dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF); Preparation of yeast cell-free extract; Radiolabeled translation products; Renilla luciferase mRNA; Room temperature; Saccharomyces Cerevisiae cells