title: Molecular cloning and immunogenicity evaluation of rotavirus structural proteins as candidate vaccine.
abstract: The middle capsid protein of rotavirus, VP6, constitutes approximately 51% of the virion by weight. The high degree of identity (>87-99%) in the primary amino acid sequences of VP6 proteins from mammalian rotaviruses suggests VP6-based vaccines could potentially provide heterotypic protection. For this reason, significant effort has been directed toward producing recombinant rotavirus VP6 vaccines. We have cloned and expressed 155bp of the VP6 most frequent in Egyptian rotavirus sewage isolates, encoding the VP6 protein in the pET30b vector having an apparent molecular mass of 12.5kDa. The recombinant VP6 expressed by the transformants was detected by SDS-PAGE analysis. Rabbits immunized with the recombinant rotavirus expressed VP6 elicited VP6-specific IgG antibodies that provided significant reductions in the infectious units of the Egyptian rotavirus sewage isolate and human reference rotavirus Wa strain in vitro assay.
authors.affiliation: Water Pollution Research Department, Environmental Research Division, National Research Centre, Dokki, Cairo, Egypt.
authors.forename: Waled M
authors.initials: WM
authors.lastname: El-Senousy
authors.forename: Yasser E
authors.initials: YE
authors.lastname: Shahein
authors.forename: Ahmed B
authors.initials: AB
authors.lastname: Barakat
authors.forename: Hossam E
authors.initials: HE
authors.lastname: Ghanem
authors.forename: Amr E
authors.initials: AE
authors.lastname: El-Hakim
authors.forename: Saeed M
authors.initials: SM
authors.lastname: Ameen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D002213:Capsid / Q000737:chemistry; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007114:Immunization; D008297:Male; D008970:Molecular Weight; D011817:Rabbits; D012401:Rotavirus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D012400:Rotavirus Infections / Q000097:blood / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D022243:Rotavirus Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D012722:Sewage / Q000382:microbiology; D014614:Vaccines, Synthetic
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D022243:Rotavirus Vaccines; D012722:Sewage; C061332:VP6 protein, Rotavirus; D014614:Vaccines, Synthetic