Characterization of messenger RNA for chick-embryo DNA polymerase beta and its translation product in vitro

Eur J Biochem. 1983 Jun 15;133(2):277-82. doi: 10.1111/j.1432-1033.1983.tb07459.x.

Abstract

A specific immunoprecipitation method, using rabbit anti-(chick DNA polymerase beta) IgG was applied to detect the polypeptide of DNA polymerase beta among translation products obtained in vitro with mRNA extracted from chick embryos. A polypeptide of Mr = 40 000 was specifically immunoprecipitated from [35S]methionine-labeled translation products and was competitive with the purified DNA polymerase beta for the antibody. Furthermore, the 40 000-Mr translation product obtained in vitro had DNA polymerase activity, which was detected by assay in situ after electrophoresis in a polyacrylamide gel containing DNA. The mRNA for DNA polymerase beta was polyadenylated and its content was estimated as the range of 0.001% of total poly(A)-rich RNA on the basis of [35S]methionine incorporation in the translation in vitro. The size of this mRNA was determined to be about 1800 nucleotides by zone sedimentation and agarose gel electrophoresis under denaturating conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Centrifugation, Density Gradient
  • Chemical Phenomena
  • Chemistry
  • Chick Embryo
  • DNA Polymerase I / genetics*
  • DNA Polymerase I / isolation & purification
  • DNA-Directed DNA Polymerase / genetics*
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Immunochemistry
  • Peptides / isolation & purification*
  • Protein Biosynthesis*
  • RNA, Messenger / isolation & purification*
  • Rabbits

Substances

  • Peptides
  • RNA, Messenger
  • DNA Polymerase I
  • DNA-Directed DNA Polymerase