Primary Screening Of Receptor Inhibitor To WSSV

In this study WSSV particles and gill cell memberane protein were purified respectively, we coated the WSSV on 96 well plate then effect with membarane protein labled with DIQ or coated the memberane protein on the plate then bound with WSSV particles; the binding result was detected by Anti-DIG-Fab-HRP system. The color change show as the index of binding activity. Also the quantities of the two composition were study. Moreover, we studied the effect on the binding by TxitionX-100, SDS and NP40. TxitionX-100 and SDS appear to higher the binding activity, otherwise NP40 have a contrary effect. By this study we established a stable method of specificly binding between WSSV and memberane receptor, which help to develope further study.In this study we searched for some drugs that maybe has function of anti-WSSV. A series of polysaccharides were investigated for their inhibiting activity towards WSSV binding. The result showed that H polysaccharides and lectins such as A polysaccharides and P polysaccharides prevented WSSV binding to memberane receptors whereas chondroitin sulfate and dextran sulfate appeared to have no affect on the binding. Moreover, the artificial infection of shrimp assay with WSSV treated by H polysaccharides were developed, and came to the similar result. The results can be illustrated that efficient infection of WSSV to target cell in initial step may require binding or adsorption to cell surface receptor of heparin sulfate(HS) proteoglycans moieties or at least that HS might serve as a primary receptor, probably concentrating WSSV in particles on the surfaces of host cell; further, this interaction is important for initiation or penetration in target cell. The date suggests that the infection of WSSV may be blocked by some source of polysaccharides.Two putative viral attachment protein (VAP) of WSSV, VP281 and VP292 were expressed successfully by yeast expressing system. The binding activity was investigated, and the result showed that VP281 had a high activity otherwise the VP282 appear to be low, which reveals that VP281 may contribute an important role to WSSV binding to target cell, and the protein expressed by yeast may retain its bioactivity. Two expressing products show strong blocking activity of WSSV binding to memberane receptors by competed and inhibit assay. Furthermore, the VP281 purified by ion change chromatography had the same result. We can further come to a conclusion that VP281 is a VAP of WSSV located on the surface of the viral membrane, while VP292 might to be. Last but not least, these show us an other fessible anti-WSSV strategy of the application of gene engineering product.