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27644505400
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Anti-HIV-1 activity was measured in lymphoid MT-4 cells by the colorimetric MTT assays as described on: M. Armand-Ugon, I. Clotet-Codina, C. Tintori, F. Manetti, B. Clotet, M. Botta, J. A. Este, Virology 2005, 343, 141-149.
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Anti-HIV-1 activity was measured in lymphoid MT-4 cells by the colorimetric MTT assays as described on: M. Armand-Ugon, I. Clotet-Codina, C. Tintori, F. Manetti, B. Clotet, M. Botta, J. A. Este, Virology 2005, 343, 141-149.
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22
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0031578901
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-1ol) and 2-4 nM RT in the presence of 8% final concentration of DMSO. All enzymes were purified to >95% purity.
-
-1ol) and 2-4 nM RT in the presence of 8% final concentration of DMSO. All enzymes were purified to >95% purity.
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23
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54849409158
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Briefly, HIV-1 RT (20-40 nM, was incubated 2 min at 37°C in a final of 4 μL in the presence of 15 nM poly(rA)/oligo(dT, 3′-OH ends, reaction buffer, and 10 mM MgCl2. Inhibitor was then added to a final of 5 μL, the concentration at which the fraction of enzyme-inhibitor complex, defined as: [E:I, E0, 1-1/ (1+[I]/Ki, was >90, Then, 145 μL of a mix containing Buffer A, 10 mM MgCl2, and 10 μM [3H]-dTTP (1 Cimm-1ol) were added at different time points. After an additional 10 min of incubation at 37°C, 50 μL aliquots were spotted on GF/C filters and acid-precipitable radioactivity measured as described for the HIV-1 RT RNA-dependent DNA polymerase activity assay.[8] The quantity representing the normalized difference between the amount of dTTP incorporated at the zero time point and at the different time points, v 0
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on.
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-
-
-
25
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54849426615
-
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i was used in the inhibition assays. Data were then plotted according to Lineweaver-Burke and Dixon.
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i was used in the inhibition assays. Data were then plotted according to Lineweaver-Burke and Dixon.
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