Cell-free translation of the messenger RNA for calf terminal deoxynucleotidyltransferase.
Poly(A)-rich RNA has been isolated from calf thymus and translated in vitro in a rabbit reticulocyte translation system. Three peptides with Mr = 58,000, 33,000, and 13,000 were specifically immunoprecipitated from the translation products with calf terminal deoxynucleotidyltransferase antiserum. An oligo(dT)-purified preparation of calf terminal transferase competed with only the Mr = 58,000 peptide in the immunoprecipitation reaction. The anti-terminal transferase serum did not precipitate a Mr = 58,000 peptide from translation products of spleen or liver mRNA, but it did precipitate the Mr = 33,000 and 13,000 peptides from products of spleen mRNA and a Mr = 13,000 peptide from products of liver mRNA. In addition, when an affinity-purified antibody to calf terminal transferase was used, only a Mr = 58,000 peptide was immunoprecipitated from the translation products of calf thymus mRNA, and none was immunoprecipitated from spleen or liver mRNA products. This antibody also precipitated a Mr = 58,000 peptide from the cell lysates of calf thymocytes labeled in vitro with [35S]methionine. These results demonstrate that calf terminal transferase is biosynthesized as a Mr = 58,000 peptide.
D000818:Animals; D000936:Antigen-Antibody Complex; D002417:Cattle; D004253:DNA Nucleotidylexotransferase / Q000235:genetics* / Q000302:isolation & purification; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D007106:Immune Sera; D008970:Molecular Weight; D011061:Poly A / Q000235:genetics; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D013950:Thymus Gland / Q000201:enzymology*
D000936:Antigen-Antibody Complex; D007106:Immune Sera; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase