【摘 要】
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AIM In order to find an effective antiviral drug against the herpes simplex virus type 1 (HSV-1) and understand its mechanism, we have screened alkalescence endonuclease (AN)-dependent inhibitors.METH
【机 构】
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School of Preclinical Medicine,Chengdu Medical College,Chengdu 610083,China;Department of Cell and M
【出 处】
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第二届世界天然药物和传统药物药理学学术会议
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AIM In order to find an effective antiviral drug against the herpes simplex virus type 1 (HSV-1) and understand its mechanism, we have screened alkalescence endonuclease (AN)-dependent inhibitors.METHODS We have constructed a plasmid,pET28-UL12, with an insertion of a DNA fragment containing the UL12 gene of HSV-1 SM44, which is 99.2% homologous to the UL12 (Gene ID: 2703382 from the GenBank), into the prokaryotic expression vector pET28a.After transformation in Escherichia coli, only kalamycin-resistant clones were selected.Expression of recombinant proteins in Escherichia coli restored the natural activity of the protein.RESULTS The anti-AN activities detected in several drugs extracted from herbs were confirmed by different degrees of inhibition on the enzymatic activities, similar to the research in Vero cells that exhibited different levels of inhibiting viral infection by these drugs.The result showed that most of these drugs can dramatically interfere with HSV-1 infection on the Veru cells, but only baicalin can significantly inhibit the rAN and stop HSV-1 replication.Docking results showed that Baicalin had a strong interaction with critical amino acid residues of AE (identity 21.12%, similar residues 41.35%, compared with rAN by DNAMAN 6.0, data not shown) which was expressed by Epstein-Barr Virus.CONCLUSION This DNA degradation assay is a useful tool to rapidly screen the inhibitors of HSV-1 at the molecular level, also representing a potential target for novel antiviral therapies.
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