Nov 1, 1977

Multihormonal induction of hepatic alpha2u-globulin mRNA as measured by hybridization to complementary DNA

Proceedings of the National Academy of Sciences of the United States of America
D T Kurtz, P Feigelson

Abstract

A procedure is presented for the preparation of a (3)H-labeled complementary DNA (cDNA) specific for the mRNA coding for alpha(2u)-globulin, a male rat liver protein under multihormonal control that represents approximately 1% of hepatic protein synthesis. Rat liver polysomes are incubated with monospecific rabbit antiserum to alpha(2u)-globulin, which binds to the nascent alpha(2u)-globulin chains on the polysomes. These antibody-polysome complexes are then adsorbed to goat antiserum to rabbit IgG that is covalently linked to p-aminobenzylcellulose. mRNA preparations are thus obtained that contain 30-40% alpha(2u)-globulin mRNA. A labeled cDNA is made to this alpha(2u)-globulin-enriched mRNA preparation by using RNA-dependent DNA polymerase (reverse transcriptase). To remove the non-alpha(2u)-globulin sequences, this cDNA preparation is hybridized to an RNA concentration x incubation time (R(0)t) of 1000 mol of ribonucleotide per liter x sec with female rat liver mRNA, which, though it shares the vast majority of mRNA sequences with male liver, contains no alpha(2u)-globulin mRNA sequences. The cDNA remaining single-stranded is isolated by hydroxylapatite chromatography and is shown to be specific for alpha(2u)-globulin mRNA b...Continue Reading

  • References14
  • Citations54
  • References14
  • Citations54

Citations

Mentioned in this Paper

Polyribosomes
Ribonucleotides
Set of Polysomal Ribosomes
Nucleic Acid Hybridization Procedure
Complex (molecular entity)
Chromatography
Immune Sera
Hepatic
Protein Biosynthesis
Globulins

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