Entry Pathway Of White Spot Syndrome Virus (WSSV) In Hematopoietic Tissue Cells, Cloning Of Clathrin Light Chain Of Cherax Quadricarinatus And Immunological Specificity Analysis Of Antibodies

White spot syndrome virus(WSSV) belongs to the Whispovirus genus of the Nimaviridae family. WSSV is a rod-shaped, non-occluded baculovirus with large doubled-stranded DNA which can infect various shrimp such as penaeid shrimp and crayfish. As a major pathogen in the shrimp farming, WSSV also has wide range of hosts in crustacean with high infection and mortality rate. However, entry pathway of WSSV in host cells are still unknown. In this thesis, entry pathway of WSSV in hematopoietic tissue(HPT) cells was analyzed by using differnet endocytic inhibitors and marker. In addition, clathrin light chain c DNA of HPT cells was cloned, and specific monoclonal antibodies were prepared. This study will not only extend our knowledge of WSSV infection, but also provide scientific evidence for prevention and control of disease in shrimp culture.(1) Entry route of WSSV in HPT cells was analyzed by using differnet endocytic inhibitors and marker. Results indicated that Chlorpromazine(CPZ), an inhibitor of clathrin-mediated endocytosis, supressed WSSV infection. Furtherly, WSSV colocalized with clathrin and transferrin, respectly, rather than cholera toxin B, which firmly confirmed that entry of WSSV depended on clathrin-mediated endocytosis. Dynamin, cholesterol and low p H were also involved in uptake of WSSV.(2) Completed sequence of clathrin light chain c DNA in Cherax quadricarinatus was obtained by cloning and RACE, designated Cq CLC(Gen Bank accession number KR075008). In phylogenetic tree analysis, evolutionary distance of clathrin light chain from C. quadricarinatus was closed to Drosophila melanogaster. Polypeptides of clathrin light chain were synthetized as antigens to immune mouses. The immunological specificity of antibodies was validated by immunofluorescence and western blot.