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Expression ánd purification of a recombinant form non-toxic Shiga toxin (Stx2) of enterohaemorrhagic Escherichia coli (EHEC)

Grant number: 13/00440-0
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): April 01, 2013
Effective date (End): March 31, 2014
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Luis Carlos de Souza Ferreira
Grantee:Mônica Josiane Rodrigues de Jesus
Home Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Stx toxin produced by strains of E. enterohaemorrhagic E. coli (EHEC), particularly by strains of serotype O157: H7 is responsible for leading sequel associated with infection, Hemolytic Uremic Syndrome (HUS), with high rates of mortality and morbidity. There are no therapies or preventive treatments against SHU but several vaccine strategies are being researched for disease control. In this project, we propose to lay the foundations for the development of a vaccine against SHU based on a recombinant form of nontoxic variant 2 of Stx (Stx2m). The work of undergraduate research will involve the establishment of a protocol optimized for the expression and purification of Stx2m from strains of E. coli BL21. The Stx2m be purified by affinity chromatography on resin containing nickel and further treated to remove the residual endotoxin. It is hoped, therefore, that the end of the project, it is possible to dispose of Stx2m able to be used in immunization trials and toxicity in vitro and in vivo, which will contribute to studies of new vaccine applications aimed at controlling the SHU.