Mutational study of sapovirus expression in insect cells
AUTOR(ES)
Hansman, Grant S
FONTE
BioMed Central
RESUMO
Human sapovirus (SaV), an agent of human gastroenteritis, cannot be grown in cell culture, but expression of the recombinant capsid protein (rVP1) in a baculovirus expression system results in the formation of virus-like particles (VLPs). In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence (Wt construct), whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent (MEG-1076 construct). While both constructs formed VLPs morphologically similar to native SaV, Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore, Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly, the position of the mutated residue was strictly conserved residue among other human SaV strains, suggesting an important role for rVP1 expression.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=553994Documentos Relacionados
- Expression of bovine vitamin K-dependent carboxylase activity in baculovirus-infected insect cells.
- Expression of processed envelope protein of hepatitis C virus in mammalian and insect cells.
- Expression of human endogenous retrovirus type K envelope glycoprotein in insect and mammalian cells.
- Rhodopsin kinase: Expression in baculovirus-infected insect cells, and characterization of post-translational modifications*
- Production of human beta interferon in insect cells infected with a baculovirus expression vector.