Open AccessPartial Purification and Characterization of the mRNA for α-Amylase from Barley Aleurone Layers
Author(s) -
Thomas J. Mozer
Publication year - 1980
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.65.5.834
Subject(s) - aleurone , messenger rna , amylase , biochemistry , chromatography , reticulocyte , lysis , centrifugation , sucrose , chemistry , fractionation , gel electrophoresis , hordeum vulgare , electrophoresis , biology , enzyme , botany , poaceae , gene
The poly(A)-containing mRNA from barley aleurone layers pretreated with gibberellic acid has been purified by phenol-chloroform extraction and repeated oligo[d(pT)]-cellulose chromatography. This RNA has been translated in both the wheat germ and reticulocyte lysate in vitro translation systems with greater than 50% of the synthesized protein being alpha-amylase. The mRNA for alpha-amylase has been further purified by dimethylsulfoxide-formamide-sucrose density gradient centrifugation and by gel electrophoresis. By these methods, its molecular weight has been determined to be 580,000.