. E.mean. Student, was St-test for comparisons between the two experiments used. A value of Po0.05 was considered statistically ABT-492 inhibitor significant. Results nuclease activity t of recombinant HSV-1 UL12 Nucleaseaktivit T of the HSV-1 UL12 was analyzed for various forms of pUC18 dsDNA and observed by agarose electrophoresis. When linear dsDNA pUC18 was treated with UL12, a smear was visible after 2 min of digestion and pUC18 dsDNA was completely Ndig degraded after 10 minutes. When was treated supercoiled pUC18 dsDNA with UL12 was initially Highest to an open circular Shaped form and then converted to the full L Length linear doppelstr Stranded DNA. With increasing incubation time, the shape was the supercoiled pUC18 dsDNA allm Hlich degraded, and the open circular Shaped and linear forms of pUC18 dsDNA were completely Degraded ndig.
These results show that recombinant HSV-1 UL12 both exonuclease and endonuclease activity Th, which are shown in accordance with earlier studies. Rheum officinale inhibits the nuclease activity of t of HSV-1 UL12 in a previous study we found that Rheum officinale, are Toosendan Paeonia suffruticosa, in Sophora flavescens Melia and capable AMG 900 945595-80-2 of HSV-1 production in Vero cells to inhibit, by preventing binding of the virus or the penetration. We are interested in whether these plants also inhibit the activity T UL12. Therefore, the methanol extracts of these plants with HSV-1 UL12 and the nuclease activity of t was mixed examined. In Figure 2, shown the methanol extract of R officinale inhibited the activity of t of a dose-UL12 Independent manner.
Three other Kr Uter showed no inhibitions on UL12 activity t. Methanol alone had no effect on UL12 activity t. Therefore, these results indicate that, additionally Tzlich to virus attachment, R. officinale exhibited an anti-UL12 activity t. Emodin inhibits the nuclease activity of t of HSV-1 UL12 with emodin specificity t is naturally present in the anthraquinone-R. officinale. That is why we are The OC, the incubation period, the SC M 1 2 3 4 5 10 20 Incubation M 1 2 3 4 5 10 20 Figure 1 nuclease activity t of the recombinant HSV-1 UL12. UL12 was with 0.1 mg of EcoRI linearized or supercoiled pUC18 dsDNA in the reaction buffer. The reaction mixtures were were grown at 37 1C for the indicated ZEITR Ume incubated, and the resulting products analyzed by electrophoresis on 1.2% agarose gel.
Lane M is the reaction in the absence of UL12 performed. Arrowheads denote the different topological forms of plasmid pUC18. OC, open circular L-shaped, linear, SC, supercoiled, HSV-1, herpes simplex virus type 1 Emodin inhibits HSV-1 yield in vitro, and TY Hsiang CY Ho British Journal of Pharmacology 229 155 227 235 curious whether the nuclease activity of emodin t inhibits HSV-1 UL12. As shown in Figure 3a, the starting DNA was completely Degraded ndig to convert in the absence of. With increasing concentrations Nukleaseaktivit t of UL12 was lead to progressively inhibited. DMSO alone had no effect on the activity of UL12-t. To further analyze the specificity t of emodin, was pUC18 dsDNA with emodin treated bovine pancreatic DNase I. mixed as shown in Figure 3b, the starting DNA was converted circular To shaped and linear forms in the presence of DNase I Open with increasing concentration, the Endonukleaseaktivit t DNase I was consistent. Therefore, these results indicate that emodin probably the active ingredient of R. officinale is that the activity of t inhibits a specific UL12. Emodin an anthraquinone compound of the three cyclic rings exists. We wonder