Cloning of protein vp1 and vp2 human parvovirus B19 in Pichia pastoris (2005)
- USP affiliated authors: DURIGON, EDISON LUIZ - ICB ; VICENTE, ELISABETE JOSE - ICB
- Unidade: ICB
- Assunto: MICROBIOLOGIA
- Language: Inglês
- Abstract: Parvovirus B19 is the only member of the Parvoviridae family known to cause disease in humans. Infection typically causes fifth disease in children, polyarthropathy syndromes in adults, transient aplastic crisis in patients with underlying chronic hemolytic anemia, and chronic anemia. Serological studies have demonstrated that parvovirus B19 IgG have a different reactivity against conformational and linear epitopes of VP1 and VP2 structural proteins, and this difference can be used as a marker of a recent or a past B19 infection. IgG reactivity toward linear epitopes on VP2 is of short duration being associated with a very recent infection, while IgG against conformational VP2 and against linear VP1 epitopes seem to persist for months or years in the majority of individuals. The objective of this work was to express the proteins VP1 and VP2 of the Human parvovirus B19 in the yeast P. pastoris that offers several advantages as: high ability of heterologous gene expression, process, folding, secretion, and simplicity of the purification steps of the expressed recombinant protein. To achieve this, the coding sequence of VP1 and VP2 were amplified by PCR from the plasmids pAE-VP1 and pAE-VP2, where these were previously cloned, and sub-cloned into the intermediary vector pGEMTeasy. These were employed in two final constructions: 1) internal expression - these two intermediary recombinant plasmids were digested with EcoR1 realizing the sequences of VP1 and VP2 that wereinserted into the expression vector pHILD2 (8,2 Kbp); 2) expression and secretion - the two intermediary recombinant plasmids were digested with EcoR1 and Not1, realizing the sequences of VP1 and VP2 that were inserted into the expression vector pPIC9K (9,3Kpb). The constructions were confirmed in the host E. coli
- Publisher: Comissão de Cultura e Extensão Universitária do ICB/USP
- Publisher place: São Paulo
- Date published: 2005
- Título do periódico: Resumos
- Conference titles: Congresso do Instituto de Ciências Biomédicas
ABNTSILVA FILHO, Claudionor Gomes da; SILVA, H. N. L.; GUERRA, O. G.; DURIGON, Edison Luiz; VICENTE, Elisabete José. Cloning of protein vp1 and vp2 human parvovirus B19 in Pichia pastoris. Anais.. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP, 2005.
APASilva Filho, C. G. da, Silva, H. N. L., Guerra, O. G., Durigon, E. L., & Vicente, E. J. (2005). Cloning of protein vp1 and vp2 human parvovirus B19 in Pichia pastoris. In Resumos. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP.
NLMSilva Filho CG da, Silva HNL, Guerra OG, Durigon EL, Vicente EJ. Cloning of protein vp1 and vp2 human parvovirus B19 in Pichia pastoris. Resumos. 2005 ;
VancouverSilva Filho CG da, Silva HNL, Guerra OG, Durigon EL, Vicente EJ. Cloning of protein vp1 and vp2 human parvovirus B19 in Pichia pastoris. Resumos. 2005 ;
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