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Cloning and expression of phospholipase A2-CB from Caudisona durissa terrificus snake venom and evaluation of its antidengue and antiyellow fever activity

Grant number: 12/12605-1
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): May 01, 2013
Effective date (End): June 30, 2017
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Victor Hugo Aquino Quintana
Grantee:Raquel Rinaldi Russo
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


Dengue virus (DENV) and yellow fever virus (YFV) belong to the genus Flavivirus, family Flaviviridae, and represent important arboviruses that cause human illness. Although a vaccine against YFV is available, many cases of yellow fever are still reported in endemic regions of the Americas and, mainly, Africa. There is no therapeutic agent to treat infection with any of these viruses. Therefore, studies to identify drugs to combat dengue and yellow fever are of great importance. Our group described recently the antiviral action of phospholipase A2 crotoxin B (PLA2-CB) from Caudisona durissa terrificus snake venom, which has action in the early stages of the replication cycle of YFV and DENV. This project aims to clone and express the two isoforms of PLA2-CB and evaluate their anti-DENV and anti-YFV activity. The sequences encoding the isoforms of the PLA2-CB will be inserted into a plasmid expression vector and will be used to transform competent E. coli cells, which will be induced to express the recombinant proteins. After confirmation and purification, the recombinants isoforms of PLA2-CB will be used in antiviral assays. The antiviral activity of the PLA2s will be evaluated in vitro by plaque-reduction test (inhibition of viral replication). The antiviral activity will be evaluated by calculation of the selectivity index (SI), which is the ratio between the cytotoxic concentration for 50% of the cell monolayer (CC50) and the concentration that inhibited 50% of the viral infection (EC50). In order to identify the sites of the PLA2-CB molecule that are directly involved in its antiviral activity, site directed mutagenesis will be carried out. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
RUSSO, RAQUEL RINALDI; DOS SANTOS JUNIOR, NILTON NASCIMENTO; OLIVEIRA CINTRA, ADELIA CRISTINA; MORAES FIGUEIREDO, LUIZ TADEU; SAMPAIO, SUELY VILELA; AQUINO, VICTOR HUGO. Expression, purification and virucidal activity of two recombinant isoforms of phospholipase A(2) from Crotalus durissus terrificus venom. ARCHIVES OF VIROLOGY, v. 164, n. 4, p. 1159-1171, . (14/02438-6, 12/12605-1)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
RUSSO, Raquel Rinaldi. Production and antiviral activity of phospholipase A2 crotoxin B recombinant isoforms. 2017. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

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Filed patent(s) as a result of this research project

PROCESSO DE OBTENÇÃO DE PROTEÍNAS RECOMBINANTES CB1-6XHIS_OPT E CB2-6XHIS_OPT, PROTEÍNAS RECOMBINANTES CB1-6XHIS_OPT E CB2-6XHIS_OPT E USO DAS PROTEÍNAS RECOMBINANTES BR1020170216276 - Universidade de São Paulo (USP) . Victor Hugo Aquino Quintana; AdeliaCristina Oliveira Cintra; Luiz Tadeu Moraes Figueiredo; Raquel Rinaldi Russo; Suely Vilela Sampaio - October 2017, 09