Dataset on recombinant expression of an ancient chitinase gene from different species of Leishmania parasites in bacteria and in Spodoptera frugiperda cells using baculovirus

Autor(es)

Cabral, Aline Diniz ^[1] ; Garcia, Felipe Baena ^[1] ; Suzuki, Rodrigo Buzinaro ^{[1, 2, 3]} ; Gois Filho, Tanil Lacerda ^[1] ; da Costa, Renata Torres ^[1] ; Pereira Vasconcelos, Ligia Marinho ^[1] ; Santos, Edmar Silva ^[1] ; Speranca, Marcia Aparecida ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Nat & Human Sci, Rua Arcturus 03, Bloco Delta, Sala 226, Lab 107, Sao Bernardo Do Campo, SP - Brazil

^[2] Marilia Med Sch, Dept Parasitol, BR-17519030 Marilia, SP - Brazil

^[3] Univ Marilia, Sch Med, BR-17525902 Marilia, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: DATA IN BRIEF; v. 32, OCT 2020.
• Citações Web of Science: 0

Resumo

The data presented here is related to negative results obtained with the recombinant expression of chitinase from four species of Leishmania parasites in two expression systems, performed in order to investigate the molecular characteristics of the Leishmania chitinase and its possible application in leishmaniasis diagnosis. Thus, heterologous Leishmania sp chitinase proteins were expressed in bacteria using the prokaryotic expression vector pET28a and Escherichia coli Mach-T1, and in Spodoptera frugiperda (Sf9) insect cells, using the eukaryotic bac-to-bac expression system (Thermo Fisher Scientific) to produce recombinant baculoviruses to infect Sf9. Biochemical and cellular analysis of the various recombinant forms of the Leishmania sp chitinase produced in prokaryotic and eukaryotic expression systems were performed through SDS-PAGE and Western blotting. Chitinase produced and purified from bacteria presented low yield and formed inactive aggregates. Heterologous chitinase obtained after infection of Sf9 insect cells with all the four Leishmania species recombinant baculoviruses presented high yield of insoluble proteins. Dot-blot serological tests presented inconclusive results against the recombinant Leishmania sp chitinases produced in both expression systems. The experiments described in this paper can help researchers to avoid errors when choosing a recombinant expression systems to produce Leishmania parasites proteins for biotechnological purposes. (C) 2020 The Author(s). Published by Elsevier Inc. (AU)

• Processo FAPESP: 13/26096-4 - Expressão heteróloga da enzima quitinase de Leishmania (L.) infantum chagasi, Leishmania (V.) braziliensis e Leishmania amazonensis: diagnóstico sorológico e estudo molecular comparativo utilizando sistemas de expressão em células de insetos e bactéria
• Beneficiário: Aline Diniz Cabral
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 16/14514-4 - Caracterização da quitinase das espécies de Leishmania endêmicas da América do Sul: utilização no diagnóstico em humanos, cães e flebotomíneos.
• Beneficiário: Marcia Aparecida Speranca
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/20221-9 - Diagnóstico molecular e sorológico das espécies de Leishmania em amostras clínicas de pacientes e cães com leishmanose visceral do centro-oeste paulista
• Beneficiário: Marcia Aparecida Speranca
• Modalidade de apoio: Auxílio à Pesquisa - Regular