Characterization of the AIDS-associated retrovirus reverse transcriptase and optimal conditions for its detection in virions.
The RNA-dependent DNA polymerase of the AIDS-associated retrovirus (ARV) gives highest activity with the synthetic template, poly(rA)oligo(dT) and prefers Mg2+ over Mn2+ as a divalent cation. It can use other template-primer combinations but with poly(rCm)oligo(dG), it prefers Mn2+ over Mg2+. Detection of ARV reverse transcriptase in culture fluids is substantially increased with an optimal KCl concentration and a special combination of EGTA and reducing agents. Our results indicate some distinguishing characteristics of the ARV reverse transcriptase and optimal conditions for its detection.
A D
AD
Hoffman
B
B
Banapour
J A
JA
Levy
D002460:Cell Line; D002478:Cells, Cultured; D017977:Deltaretrovirus / Q000201:enzymology* / Q000254:growth & development; D004533:Egtazic Acid / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009000:Monocytes / Q000201:enzymology / Q000382:microbiology; D017830:Octoxynol; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013698:Templates, Genetic; D014771:Virion / Q000201:enzymology
D009838:Oligodeoxyribonucleotides; D013439:Sulfhydryl Reagents; D011061:Poly A; D011092:Polyethylene Glycols; D008345:Manganese; D004533:Egtazic Acid; C018368:poly(rA).oligo(dT); D011189:Potassium Chloride; D017830:Octoxynol; D012194:RNA-Directed DNA Polymerase; D008274:Magnesium