Expression and characterization of Sapporo-like human calicivirus capsid proteins in baculovirus.
Sapporo-like caliciviruses reveal typical calicivirus morphology and cause acute gastroenteritis. This study describes the expression in baculovirus of capsid proteins of two Sapporo-like calicivirus strains (Hou/86 and Hou/90). Eight different constructs of the capsid genes were compared for production of the proteins. Constructs containing short (9 or 19 nt) upstream sequences failed to produce capsid proteins but extension of the upstream sequence to 73 nt resulted in production of capsid proteins. Expressed capsid protein with the MEG tri-peptide as the N-terminus self-formed virus-like particles (VLPs). Expressed protein with an upstream AUG failed to form VLPs. Addition of His-tag to the N-terminus of capsid protein also blocked VLP formation. Of three Norwalk-Hou/90 chimeric capsid gene constructs, one resulted in production of chimeric capsid and the protein did not form VLPs. Recombinant capsid proteins for each of Hou/86 and Hou/90 were further characterized. The expressed capsid antigens of the two strains were antigenically distinct but shared a common epitope(s). Further study of these proteins should allow development of immunologic assays for diagnosis and should help to clarify the epidemiology of Sapporo-like caliciviruses in humans.
Jiang
X
X
Center for Pediatric Research, Eastern Virginia Medical School, Children's Hospital of The King's Daughters, Norfolk 23510, USA. jjiang@chkd.com
Zhong
W
W
Kaplan
M
M
Pickering
L K
LK
Matson
D O
DO
D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D002139:Caliciviridae / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D017250:Caliciviridae Infections / Q000175:diagnosis* / Q000821:virology; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D002648:Child; D004196:Disease Outbreaks; D000939:Epitopes / Q000276:immunology; D005759:Gastroenteritis / Q000175:diagnosis / Q000821:virology; D006801:Humans; D007124:Immunoenzyme Techniques; D011994:Recombinant Proteins / Q000096:biosynthesis; D018411:Spodoptera; D014771:Virion / Q000378:metabolism / Q000648:ultrastructure
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D011994:Recombinant Proteins