Reverse transcription of thyroglobulin 33-S mRNA.
Bovine thyroglobulin 33-S mRNA has been used as a template for the synthesis of a complementary DNA, using RNA-directed DNA polymerase from the avian myeloblastosis virus. The yield of the reaction was relatively poor and the size of the cDNA did not exceed 10 S. Nevertheless, a copy of high specific radioactivity (approximately 10(7) counts. min-1 microgram-1) could be obtained which hybridized specifically back to its template with an rot1/2 value about 5 times higher than that observed in hybridizations between hemoglobin mRNA (alpha + beta chain) and hemoglobin cDNA. This suggests that thyroglobulin mRNA does not contain extensive internal repetitive sequences. Quantification of thyroglobulin mRNA sequences among various RNA preparations from the beef thyroid was performed using cDNA/RNA hybridizations in RNA excess. The results confirmed that thyroglobulin mRNA represents the large majority of mRNA in membrane-bound polysomes and indicated the virtual absence of thyroglobulin sequences on free polyosomes. The cDNA transcribed from mRNA of bovine origin hybridized efficiently with thyroid RNA from goats, dogs and humans. Although the heterologuous hybrids exhibited the expected decrease in thermal stability, the bovine cDNA provides an appropriate probe for studies dealing with the expression of the thyroglobulin gene in various mammals including man.
D000818:Animals; D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D002417:Cattle; D004247:DNA; D004285:Dogs; D006041:Goats; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D011061:Poly A; D012333:RNA, Messenger / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013698:Templates, Genetic; D013954:Thyroglobulin / Q000096:biosynthesis*; D013961:Thyroid Gland / Q000378:metabolism*
D012333:RNA, Messenger; D011061:Poly A; D004247:DNA; D013954:Thyroglobulin; D012194:RNA-Directed DNA Polymerase