Reverse transcriptase activity per virion for avian myeloblastosis virus and Rauscher murine leukemia virus.
We have measured reverse transcriptase enzyme activity per virus particle for samples of avian myeloblastosis virus (BAI strain) and murine leukemia virus (RAUSCHER) USing the synthetic template poly(rC)-oligo(dG). Absolute virus concentrations were determined directly by laser beat frequency spectroscopy. Enzyme activity per virion was determined from the slope of the activity plotted as a function of virus concentration. With this reverse transcriptase assay, the minimum activity (expressed as picomoles of dGTP incorporated/virion per hour) is estimated at (28.1 +/- 4.2) X 10(-7) for avian myeloblastosis virus and (1.1 +/- 0.2) X 10(-7) for murine leukemia virus. The sensitivity of this assay, which is determined by the level of incorporated radioactivity measurable above background, is 2.5 X 10(-4) virions for avian myeloblastosis virus (with dGTP specific activity of 8.9 Ci/mmol) and 88 X 10(-4) virions for murine leukemia virus (with dGTP specific activity of 6.52 CI/mmol). These results show that although reverse transcriptase assays can obviously be used to measure relative virus concentrations of equally purified samples of the same virus, they can be very misleading when used to compare the concentrations of different virus species.
D001354:Avian Leukosis Virus / Q000201:enzymology*; D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D002474:Cell-Free System; D004279:DNA, Viral / Q000096:biosynthesis; D007834:Lasers; D011119:Polynucleotides / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011925:Rauscher Virus / Q000201:enzymology*; D012542:Scattering, Radiation; D013057:Spectrum Analysis; D013698:Templates, Genetic
D004279:DNA, Viral; D011119:Polynucleotides; D012194:RNA-Directed DNA Polymerase