Title:	Desenvolvimento de vetores virais para produção de proteínas recombinantes
Authors:	Bonnet, Raquel Medeiros Vasques
Orientador(es)::	Nagata, Tatsuya
Assunto::	Vetor viral Biofármacos Proteínas recombinantes Vírus vegetais
Data de defesa::	26-Oct-2018
Citation:	BONNET, Raquel Medeiros Vasques. Desenvolvimento de vetores virais para produção de proteínas recombinantes. 2018. 110 f., il. Tese (Doutorado em Biotecnologia e Biodiversidade)—Programa em Rede Multi-Institucional do Pró-Centro-Oeste de Pós-Graduação em Biotecnologia e Biodiversidade, Universidade de Brasília, Universidade Federal de Goiás, Universidade Federal do Mato Grosso, Universidade Federal do Mato Grosso do Sul, Universidade Federal da Grande Dourados, Brasília, 2018.
Abstract:	The use of plant viral vectors for transient expression of heterologous proteins is a useful tool for large-scale production of important biopharmaceuticals such as antibodies and antigens. Some groups of plant viruses are widely used for this purpose, for example tobamovirus, potexvirus, cucumovirus, and so on. In this study, three different viral vectors were developed. The tobamovirus Pepper mild mottle virus (PMMoV) was used for the development of the first viral vector in this study for the expression of recombinant proteins. Initially, the PMMoV infectious clone was constructed on the small binary vector pJL89, replacing a middle part of the viral coat protein gene by the Green fluorescent protein (GFP) reporter gene, using the Gibson Assembly technique. The fused pJL_PMMoV(Δ)GFP vector proved to be viable for protein expression. The GFP gene was replaced by the viral antigen gene of Chikungunya virus (CHIKV) and successfully expressed. In the second work, the tymovirus infectious clone Tomato blistering mosaic virus – ToBMV was adapted to be used as display peptide vector, in the future. The N-terminal region of the protein coat was modified by deleting 24 amino acids (CPΔ2-24), which were shown to be non-essential for expression of the capsomer and infectivity. And, finally in the third study, a recombinant baculovirus was constructed to check a set of tymovirus-like particles (tVLPs) following expression of the wild-type CP or a version containing a small deletion in the amino-terminal region of CP (CPΔ2-24). To this region was added a small sequence containing CHIKV epitope to test the potential use of ToBMV as a protein carrier of immunogenic epitopes. Likewise, cell formation was observed in all versions of CP when the gene was expressed in insect cells by baculovirus.
metadata.dc.description.unidade:	Instituto de Ciências Biológicas (IB)
Description:	Tese (doutorado)—Universidade de Brasília, Universidade Federal de Goiás, Universidade Federal do Mato Grosso, Universidade Federal do Mato Grosso do Sul, Universidade Federal da Grande Dourados— Programa em Rede Multi-Institucional do Pró-Centro-Oeste de Pós-Graduação em Biotecnologia e Biodiversidade, 2018.
metadata.dc.description.ppg:	Programa de Pós-Graduação em Biotecnologia e Biodiversidade (Rede PRÓ-CENTRO-OESTE)
Agência financiadora:	Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
Appears in Collections:	Teses, dissertações e produtos pós-doutorado

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