Expression and self-assembly of Grimsby virus: antigenic distinction from Norwalk and Mexico viruses.
A cDNA obtained from Grimsby virus (GRV), a Norwalk-like virus, purified from a stool sample of a symptomatic adult associated with a gastroenteritis outbreak in the United Kingdom, was used to obtain the complete nucleotide sequence of the second open reading frame (ORF2). The ORF2 sequence of GRV predicts a capsid of 539 amino acids (aa) which exhibits aa identities of 96% to Lordsdale virus, 67% to Mexico virus (MXV), and 43% to Norwalk virus (NV). The GRV capsid protein was expressed in insects cells by using a recombinant baculovirus, and the resulting virus-like particles (VLPs) possessed a protein with an apparent molecular weight of 58,000. Hyperimmune antisera raised against purified GRV, MXV, and NV VLPs were tested in an indirect enzyme-linked immunosorbent assay (ELISA) against GRV, NV, and MXV VLPs, revealing that GRV is antigenically distinct from both NV and MXV. The antigenic specificity of the GRV-hyperimmune antiserum was confirmed in an antigen capture ELISA using GRV-, NV-, or MXV-containing fecal specimens. The expression of the GRV capsid protein has, for the first time, allowed the antigenic comparison of three distinct recombinant Norwalk-like viruses.
Hale
A D
AD
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA.
Crawford
S E
SE
Ciarlet
M
M
Green
J
J
Gallimore
C
C
Brown
D W
DW
Jiang
X
X
Estes
M K
MK
D000328:Adult; D000595:Amino Acid Sequence; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics*; D002139:Caliciviridae / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D017250:Caliciviridae Infections / Q000821:virology; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005759:Gastroenteritis / Q000821:virology; D015870:Gene Expression; D005814:Genes, Viral; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009663:Norwalk virus / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D016366:Open Reading Frames; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral; D018076:DNA, Complementary; D004279:DNA, Viral