Poster (Painel)
559-1Identification, molecular cloning and expression analysis of the elongation factor Tu of Paracoccidioides brasiliensis.
Autores:Caroline Maria Marcos (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Haroldo Cesar de Oliveira (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Julhiany de Fátima da Silva (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Patricia Akemi Assato (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Maria José Soares Mendes Giannini (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara) ; Ana Marisa Fusco Almeida (FCF-UNESP - Faculdade de Ciências Farmacêuticas de Araraquara)


Paracoccidioidomycosis (PCM) is a disease caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb) and is highly prevalent in Brazil, where it is the principal cause of death by systemic mycoses. Most pathogens to express surface factors that mediate binding to host cells, in this case, indirectly, the binding to host adhesion components such as extracellular matrix (ECM) or proteins, which act as "interlinking" molecules. The ECM plays an important role in regulation of cell adhesion, differentiation, migration and proliferation of cells. Our group is involved in the study of proteins related to adhesion and invasion of Pb to eukaryotic cells. Then, a protein preferentially expressed in yeast cells cultured in BHI medium supplemented with 5% sheep blood with molecular mass of 44 kDa was isolated from the proteome and identified by mass spectrometry as an elongation factor Tu (EF-Tu). EF-Tu, conserved in most organisms, is a member of a still expanding family of “moonlighting proteins” located in other pathogens in the cytoplasm and at the cell surface and was identified as a Factor H, fibronectin and plasminogen binding protein can then facilitate the adhesion and invasion of host tissue as well evasion of the complement system. So far there are no reports on its role in pathogenic fungi. The main objective of this study was to clone, identify and to express this protein. After the amplification of 1800bp fragment, related to Pb EF-Tu gene, was cloned into the expression vector pET TOPO. E. coli BL21 Star® were transformed with the recombinant vector and the expression of EF-Tu recombinant protein was induced by adding 0,4mM of IPTG in liquid cultures. After induction, a protein EF-Tu has been detected by SDS-PAGE. We believe this can be a potential target for the design of new drugs for the prevention of fungal adhesive process.

Palavras-chave:  Paracoccidioides brasiliensis, elongation factor Tu, recombinant protein