Deoxyribonucleic Acid Polymerase of Rous Sarcoma Virus: Reaction Conditions and Analysis of the Reaction Product Nucleic Acids
AUTOR(ES)
Bishop, D. H. L.
RESUMO
Reaction conditions for Rous sarcoma virus ribonucleic acid (RNA)-instructed deoxyribonucleic acid (DNA) polymerase activity are described whereby the viral RNA is relatively protected from endogenous or added nuclease activity. Three analyses of reaction product nucleic acids (3H-RNA, 32P-DNA) were compared, namely, gel electrophoresis, Cs2SO4 gradient centrifugation, and hydroxyapatite column chromatography. It was found that hydroxyapatite analysis could be misleading unless the state of the template RNA was monitored concomitantly with the DNA analysis. Gel electrophoresis and Cs2SO4 gradient centrifugation gave comparable results. It was concluded that analyses of the product of reverse transcriptase reactions should not only refer to the template RNA and product DNA species, but also be performed with virus or viral RNA which do not have or obtain nicks in the 60S RNA. Otherwise, interpretation of the results would have the ambiguity of potential artifacts caused by those degraded RNA molecules.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=376254Documentos Relacionados
- Deoxyribonucleic Acid Polymerase Associated with Rous Sarcoma Virus and Avian Myeloblastosis Virus: Properties of the Enzyme and Its Product
- Deoxyribonucleic Acid Polymerase(s) of Rous Sarcoma Virus: Effects of Virion-Associated Endonuclease on the Enzymatic Product
- Hybridization of Rous Sarcoma Virus Deoxyribonucleic Acid Polymerase Product and Ribonucleic Acids from Chicken and Rat Cells Infected with Rous Sarcoma Virus 1
- Deoxyribonucleic Acid Polymerase of Rous Sarcoma Virus: Studies on the Mechanism of Double-Stranded Deoxyribonucleic Acid Synthesis
- Deoxyribonucleic Acid Polymerases of Rous Sarcoma Virus: Kinetics of Deoxyribonucleic Acid Synthesis and Specificity of the Products
