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Characterization of the cDNA synthesized by avian retrovirus reverse transcriptase using 35 S avian myeloblastosis virus RNA and an exogenous bovine primer tRNA
Author(s) -
Sarih Leila,
Araya Alejandro,
Litvak Simon
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80642-2
Subject(s) - reverse transcriptase , complementary dna , primer (cosmetics) , microbiology and biotechnology , rna , biology , primer binding site , rna directed dna polymerase , retrovirus , transfer rna , polymerase , dna , virus , biochemistry , chemistry , virology , gene , organic chemistry
Bovine tRNA Trp can be partially hybridized to the avian myeloblastosis virus (AMV) 35 S RNA at 37°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.