Of Wssv Infection Of Shrimp Blood Cells And Its Binding Protein In The Blood Cell Membrane

Primary haemocyte from heart of Penaeus chinensis were cultured by use of leibovitz's 2 XL-15 medium consisting of 20% FBS and other supplements, adjusting osmolarity with sodium chloride, pH 1.2-1 A, and at a temperature of 29C.The haemocyte remained viable for about 20 days. Cells were inoculated with WSSV. The infection was confirmed with immunocytochemistry (ICC) test. It revealed that WSSV had entered into the cell nucleolus. It was detected that WSSV had little propagated in infected cells in different times with same virus dilution and in the same times with different virus concentration, cells were also inoculated with mixture of WSSV -and it's monoclonal antibodies, the results showed that the neutralizing effects of each monoclonal antibodies were different, and the monoclonal antibodies 6B4, 2D2 were best.Penaeus chinensis hemocyte are permissive towards infection with WSSV. In order to detect receptors on membrane of Penaeus chinensis hemocyte, proteins of intact Penaeus chinensis hemocyte were tested in virus overlay protein binding assay (VOPBAs), three WSSV binding protein 83.0, 74.0, 69.9kDa were revealed. The cell membrane from Penaeus chinensis hemocyte was prepared and used for sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Two membrane protein 74.0, 69.9kDa that were proved to be virus binding protein were found. The characteristics of these proteins remain to be studied further. Furthermore, the hemocyte of five crustaceas, Penaeus chinensis , Penaeus vannamei, Penaeus japonicus, Squilla and Crab were prepared inorder to compared their protein difference.