Both probes were synthesized through PCR by using KlenTaq and P.F.U. polymerases, vector-specific primers, 100 jJuCi [a-32P]dATP, 20 |xM dATP, and 200 |xM each of deoxycytidine triphosphate (dCTP), deoxyguanidine triphosphate (dGTP), and deoxythymidine triphosphate (dTTP). Both hybridizations were performed at 42°C for 16 h, and the membrane was washed and exposed to XAR film at -80°C in the presence of intensifying screens.
To determine antiproliferative activity, the concentration of ovIFN-т required to inhibit cell proliferation by 50% of that of controls was determined by linear regression analysis. Differences in both antiviral and antiproliferative activity between ovIFN-ts were determined by least-squares analysis of variance (LS-ANOVA) after data had been log-transformed. buy flovent inhaler
Life span of the CL was defined in days from the time of the synchronized estrus to the subsequent fall in serum progesterone (< 0.5 ng/ml). Ewes that were not determined to be in estrus by Day 45 postestrus were killed and were assigned a CL life span of 45 days. Values were transformed by nonparametric ranking and analyzed by LS-ANOVA. Differences in treatment means were partitioned further by completing orthogonal contrasts (60 vs. 300 |JLg/day p3; 60 vs. 300 |xg/day рЗТгпб; 60 and 300 (jLg/day p3 vs. 60 and 300 (xg/day рЗТгпб; control vs. all other treatments).