Proteomic Analysis of Cytoskeleton Proteins in Fish.
In this chapter, we describe laboratory protocols for rearing fish and a simple and efficient method of extracting and identifying pathogen and host proteins that may be involved in entry and replication of commercially important fish viruses. We have used the common carp (Cyprinus carpio L.) and goldfish (Cyprinus auratus) as a model system for studies of proteins involved in viral entry and replication. The chapter describes detailed protocols for maintenance of carp, cell culture, antibody purification of proteins, and use of electrospray-ionization mass spectrometry analysis to screen and identify cytoskeleton and other proteins that may be involved in viral infection and propagation in fish.
Department of Biology, Technion - Israel Institute of Technology, Technion, Haifa, Israel.
Michael
M
Gotesman
Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Veterinärplatz 1, Vienna, 1210, Austria.
Simon
S
Menanteau-Ledouble
Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Veterinärplatz 1, Vienna, 1210, Austria. matboulim@vetmeduni.ac.at.
Mansour
M
El-Matbouli
D018138:Alphaherpesvirinae / Q000502:physiology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002347:Carps / Q000378:metabolism* / Q000821:virology; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000276:immunology / Q000378:metabolism*; D029941:Fish Proteins / Q000276:immunology / Q000378:metabolism*; D006054:Goldfish / Q000378:metabolism* / Q000821:virology; D007267:Injections; D018096:Mononegavirales / Q000502:physiology; D040901:Proteomics / Q000379:methods*; D053586:Virus Internalization; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D003598:Cytoskeletal Proteins; D029941:Fish Proteins
Carp; Cyprinid herpesvirus 3 (CyHV-3); Cyprinidae fish; Electrospray-ionization mass spectrometry (ESI-MS) analysis; Spring viremia of carp virus (SVCV)