title: Identification of two epitopes in the carboxyterminal 15 amino acids of the E1 glycoprotein of mouse hepatitis virus A59 by using hybrid proteins.
abstract: cDNA fragments coding for the carboxy terminus of the E1 envelope glycoprotein from mouse hepatitis virus A59, a coronavirus, were cloned into the bacterial expression vector pEX. Clones expressing E1 antigenic determinants were selected with a polyclonal anti-E1 antibody and used for immunization of rabbits and for affinity purification of existing polyclonal antisera. Immunofluorescence testing and immunoperoxidase labeling of coronavirus-infected cells showed that these reagents were monospecific for E1. In addition, by using hybrid proteins containing different lengths of the E1 carboxy terminus to affinity-purify a polyclonal antiserum against E1, we have been able to define two epitopes within the last 15 amino acid residues of the protein. These epitope-specific antibodies bind to E1 in Golgi and perinuclear membranes as well as to budding viruses; they do not, however, label the plasma membrane or the membranes of post-Golgi vesicles transporting virions to the cell surface.
mesh_terms: D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000276:immunology*; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000276:immunology; D000939:Epitopes; D006023:Glycoproteins / Q000235:genetics; D007425:Intracellular Membranes / Q000648:ultrastructure; D008565:Membrane Proteins / Q000276:immunology; D008970:Molecular Weight; D006517:Murine hepatitis virus / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins