coli DH10Bac for the construction of recombinant bacmids. These bacmids, containing the sequence of the protein with antiviral activity and other chosen proteins, were used for the expression of the proteins in a baculovirus/Sf9 cells system. Three passages of the recombinant virus were performed in Sf9 cell cultures so far. At the moment, titers of the baculovirus obtained

in the different passages as well as the antiviral activity of the recombinant protein produced in this system were determined. To eliminate the possibility that the observed effect is due to characteristics of the www.selleckchem.com/products/azd5363.html construct other than the antiviral activity itself, we used the same approach and procedures to construct recombinant bacmids expressing other L. obliqua proteins, namely LOH-19 and 8-LOH ( Veiga et al., 2005). These two recombinant bacmids, as well as an empty bacmid were used Selleck AZD0530 to treat Sf-9 cells infected with a picornavirus. The results showed that the empty bacmid or those expressing the other recombinant proteins were not effective in inhibiting the replication of EMC

virus, presenting results similar to those of the control of infected cells and of the untreated cells. On the other hand, when infected cultures were treated with the recombinant antiviral, there was a reduction of about 3 logs in the viral titers in comparison to that of controls. Therefore, when the purified antiviral protein was used, the reduction in virus produced was around 4 logs, showing that the recombinant antiviral protein remained fully

active ( Table 1). We are currently testing the effect of the antiviral purified recombinant protein on enveloped viruses (measles, rubella and herpes simplex). Preliminary data have shown that the purified recombinant protein is able to reduce by at least 4 logs the replication of the rubella virus and by about 6 logs the replication of the herpes simplex virus (data not shown). To facilitate purification, a His-tag sequence was included in the C-terminal region of the proteins rAVLO, LOH-19-AY829833 and 8-LOH. The protein was separated by SDS–PAGE and transferred to nitrocellulose membranes (Sambrook and Russell, 2001). After transfer, the membrane was marked with the anti-histidine antibody to confirm the presence of the protein. The result is shown in Fig. 3. As can be seen, there was the presence of a band with Idoxuridine strong labeling with the antibody, demonstrating the expression of the antiviral protein. Viral diseases affect hundreds of millions of people worldwide every year. Even though some antiviral drugs are under clinical trials, 50% of them are directed toward the treatment of HIV. Therefore, there is a need for the development of antiviral agents specific for emerging newly-recognized human pathogens (such as SARS coronavirus and influenza viruses H5N1 and H1N1) (Delcroix and Riley, 2011). Recently, various studies have reported the antiviral properties in products obtained from arthropods.