Characterization of the cDNA synthesized by avian retrovirus reverse transcriptase using 35 S avian myeloblastosis virus RNA and an exogenous bovine primer tRNA.
Bovine tRNA(Trp) can be partially hybridized to the avian myeloblastosis virus (AMV) 35 S RNA at 37 degrees C, in the presence of AMV RNA-dependent DNA polymerase (reverse transcriptase). This template-primer complex is active in the synthesis of viral cDNA. The size of the cDNA products synthesized in the in vitro reconstituted AMV system was determined by urea-polyacrylamide gel electrophoresis using a tRNA labelled at the 3'-end by yeast tRNA nucleotidyl transferase. The synthesized cDNA has a size of about 100 nucleotides and was shown by Southern blotting to be complementary to a specific sequence of the 5'-end of the retroviral genome. These results indicate that reverse transcriptase is able to anneal the exogenous primer tRNA at the 'primer-binding site' near the 5'-end of the long terminal repeat (LTR) of AMV RNA.
D001354:Avian Leukosis Virus / Q000201:enzymology / Q000235:genetics; D009189:Avian Myeloblastosis Virus / Q000201:enzymology / Q000235:genetics; D003101:Collodion; D004247:DNA / Q000096:biosynthesis*; D004262:DNA Restriction Enzymes; D009693:Nucleic Acid Hybridization; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012364:RNA, Transfer, Trp / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013698:Templates, Genetic
D012345:RNA, Transfer, Amino Acid-Specific; D012364:RNA, Transfer, Trp; D012367:RNA, Viral; D003101:Collodion; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004262:DNA Restriction Enzymes