Expression and purification of structural proteins pr and prM from flavivirus.

• Grant number: 23/14340-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2023
• End date: March 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Aparecida Sadae Tanaka
• Grantee: Alice Mesquita Gonçalves
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 19/03779-5 - Use of phage display as a tool in the diagnosis and control of diseases transmitted by hematophagous vectors, AP.TEM

Abstract

Hematophagous insects, such as the Aedes aegypti mosquito, are responsible for the spread of neglected diseases and consequently, countless deaths throughout the world. Ae. aegypti is the main vector of viruses in Brazil, including dengue, Chikungunya, yellow fever and Zika. To date, the only vaccine available is for yellow fever and there are no vaccines or treatments available for other viruses. Controlling the transmission of these viruses is done by controlling the vector, Ae. aegypti, mainly with the use of insecticides. In addition to these problems, the identification of viruses circulating during an outbreak is also not fast and specific, as the protein of these viruses are highly similar. In an attempt to contribute knowledge in this area, the present project aims are: express, purify and characterize non-structural proteins (pr and prM) of the dengue 2, dengue 4 and zika viruses. The proteins will be expressed in bacteria, then solubilized and purified by Ni-Sepharose affinity chromatography and if necessary, they will be subjected to other chromatographic methods. The purified proteins will be quantified by the Bradford method and analysed by SDS-PAGE. The purified proteins will be used in the future to select specific recombinant antibodies. In parallel with the production of proteins, this project will also provide learning and technical training for the student, as well as lessons from experience in a research laboratory.