title: Comparison of two eukaryotic systems for the expression of VP6 protein of rotavirus specie A: transient gene expression in HEK293-T cells and insect cell-baculovirus system.
abstract: The VP6 protein of rotavirus A (RVA) is a target antigen used for diagnostic assays and also for the development of new RVA vaccines. We have compared the expression of VP6 protein in human embryonic kidney (HEK293-T) cells with results obtained using a well-established insect cell-baculovirus system. The recombinant VP6 (rVP6) expressed in HEK293-T cells did not present degradation and also retained the ability to form trimers. In the insect cell-baculovirus system, rVP6 was expressed at higher levels and with protein degradation as well as partial loss of ability to form trimers was observed. Therefore, HEK293-T cells represent a less laborious alternative system than insect cells for expression of rVP6 from human RVA.
authors.affiliation: Laboratório de Virologia Comparada e Ambiental, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz. Av. Brasil, 4365, Pavilhão Hélio & Peggy Pereira, Rio de Janeiro, RJ 21040-360, Brazil. haroldo.cid@bio.fiocruz.br
authors.forename: Haroldo Cid
authors.initials: HC
authors.lastname: da Silva Junior
authors.forename: Sérgio
authors.initials: S
authors.lastname: da Silva E Mouta Junior
authors.forename: Marcos César Lima
authors.initials: MC
authors.lastname: de Mendonça
authors.forename: Mirian Claudia
authors.initials: MC
authors.lastname: de Souza Pereira
authors.forename: Alanderson
authors.initials: A
authors.lastname: da Rocha Nogueira
authors.forename: Maria Luiza Borges
authors.initials: ML
authors.lastname: de Azevedo
authors.forename: José Paulo Gagliardi
authors.initials: JP
authors.lastname: Leite
authors.forename: Márcia Terezinha Baroni
authors.initials: MT
authors.lastname: de Moraes
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology / Q000379:methods*; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D007313:Insecta; D059748:Proteolysis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012401:Rotavirus / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C061332:VP6 protein, Rotavirus