Cloning and expression of FECV spike gene in vaccinia virus. Immunization with FECV S causes early death after FIPV challenge.
The spike gene of the feline enteric coronavirus (FECV), strain FECV-1683, was PCR amplified from total RNA extracted from FECV-infected cells and its sequence determined. A primary translation product of 1454 amino acids is predicted from the nucleotide sequence, containing a N-terminal signal sequence, a C-terminal transmembrane region and 33 potential N-glycosylation sites. The sequence shares 92% homology with the previously published feline infectious peritonitis virus, strain WSU-1146; however, several regions were identified that distinguished FECV from Feline Infectious Peritonitis virus, FIPV. The full length FECV S gene was cloned and expressed in vaccinia virus. Recombinants produced a 200 kD protein which was recognized by sera from cats infected with FIPV. When kittens were immunized with the vaccinia/FECV S recombinant, neutralizing antibodies to FIPV were induced. After challenge with a lethal dose of FIPV, the recombinant vaccinated animals died earlier than control animals immunized with vaccinia virus alone.
Klepfer
S
S
Department of Molecular Biology, Smithkline Beecham Animal Health, King of Prussia, Pennsylvania, USA.
Reed
A P
AP
Martinez
M
M
Bhogal
B
B
Jones
E
E
Miller
T J
TJ
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000097:blood; D000917:Antibody Formation; D015153:Blotting, Western; D002415:Cats; D002460:Cell Line; D003001:Cloning, Molecular; D016765:Coronavirus, Feline / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016766:Feline Infectious Peritonitis / Q000276:immunology* / Q000503:physiopathology; D005814:Genes, Viral; D007114:Immunization; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology*; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000096:biosynthesis; D012367:RNA, Viral / Q000302:isolation & purification; D064370:Spike Glycoprotein, Coronavirus; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000276:immunology*
D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals; D012367:RNA, Viral; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins