Characterization of messenger RNA for chick-embryo DNA polymerase beta and its translation product in vitro.
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.
D000818:Animals; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D002642:Chick Embryo; D004256:DNA Polymerase I / Q000235:genetics* / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007120:Immunochemistry; D010455:Peptides / Q000302:isolation & purification*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification*; D011817:Rabbits
D010455:Peptides; D012333:RNA, Messenger; D004256:DNA Polymerase I; D004259:DNA-Directed DNA Polymerase