Expressão heteróloga da proteína não-estrutural 1 (NS1) do vírus dengue-2 em Arabidopsis thaliana

A dengue é a doença mais importante causada por um arbovírus no mundo. O vírus da dengue pertence à família Flaviviridae e possui 4 sorotipos antigenicamente distintos. São vírus de RNA fita simples, polaridade positiva, com o genoma de aproximadamente 11 Kb que é traduzido em uma poliproteína subdi...

Nível de Acesso:openAccess
Publication Date:2015
Main Author: Xisto, Mariana Fonseca lattes
Orientador/a: Paula, Sérgio Oliveira de
Format: Dissertação
Language:por
Published: Universidade Federal de Viçosa
Áreas de Conhecimento:
Online Access:http://www.locus.ufv.br/handle/123456789/6696
Citação:XISTO, Mariana Fonseca. Expressão heteróloga da proteína não-estrutural 1 (NS1) do vírus dengue-2 em Arabidopsis thaliana. ano. 2015 f. Dissertação (Mestrado em Biologia Celular e Estrutural) - Universidade Federal de Viçosa, Viçosa. 2015.
Resumo inglês:Dengue fever is the most important disease caused by an arbovirus in the world. The dengue virus belongs to the Flaviviridae family and has four antigenically distinct serotypes. They are single stranded RNA viruses, positive polarity, with a genome of approximately 11 Kb which is translated into a polyprotein subdivided in three structural proteins and seven non-structural proteins (that are related to viral replication, expression of viral proteins and virulence of serotypes). In the last twenty years, it was observed a significant increase in the epidemic activity, expansion of the geographical distribution and streaming of different serotypes in areas where the disease was not prevalent. One of the most alarming events was the increase in the number of cases of dengue hemorrhagic fever (DHF) in the Americas. A further limiting point of the diagnostic tests that capture antibody present in the serum of infected patients is the difficulty of obtaining low-cost and large-scale production of the antigens. Due to this factor, we aimed to build a system in transgenic Arabidopsis thaliana expressing the nonstructural protein (NS1) of the dengue-2 virus. The gene of the NS1 protein was optimized for expression in plants and cloned into the plasmid pCAMBIA3301. The expression vector pCAMBIA3301/NS1 was used for transformation of Arabidopsis thaliana. The third generation of transformed plants were selected by dominant homozygous. The construction of a transgenic Arabidopsis thaliana system was demonstrated by immunolocalization of the NS1 protein into the lumen of the endoplasmic reticulum in the leaf tissues. This system will be used to obtain the NS1 antigen with application purpose of dengue diagnostic tests.