Processing and antigenicity of entire and anchor-free spike glycoprotein S of coronavirus TGEV expressed by recombinant baculovirus.
The S gene of transmissible gastroenteritis virus (TGEV) was inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV) using the transfer plasmid pVL941. Infection of Sf9 insect cells with the recombinant virus resulted in the synthesis of a 175K polypeptide which was able to trimerize and was transported to the cell surface as is the authentic TGEV S protein. Despite the lack of complete carbohydrate processing, the recombinant S protein exhibited antigenic properties similar to TGEV S and induced high levels of neutralizing antibodies in immunized rats. Engineering a deletion (70 amino acids) into the carboxy-terminus containing the membrane anchor of the polypeptide allowed its secretion. The oligomerization process and the antigenic profile of the anchor-free S protein were shown to be partially altered.
Godet
M
M
Laboratoire de Virologie et Immunologie Moléculaires, Institute National de la Recherche Agronomique, Jouy-en-Josas, France.
Rasschaert
D
D
Laude
H
H
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D002498:Centrifugation; D015870:Gene Expression / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005760:Transmissible gastroenteritis virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D006023:Glycoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C064484:spike protein S, Transmissible gastroenteritis virus