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Prep Biochem Biotechnol ; 48(10): 968-976, 2018.
Article in English | MEDLINE | ID: mdl-30489218

ABSTRACT

Leishmaniosis is a complex of diseases that can be fatal, if not given proper attention. Despite its relevance in the public health system, there is no vaccine capable of preventing the disease in humans so far and its treatment is expensive and aggressive to human health. The present study aims to optimize the induction parameters of the 503 Leishmania i. chagasi antigen expressed in recombinant Escherichia coli M15. The induction at different cell densities was evaluated in order to analyze the influence of the induction time on the yield of the protein of interest. In this segment, lactose and isopropyl-ß-d-thiogalactopyranoside (IPTG) were used as inducer molecules, using various concentrations: 0.1 g/L, 1.0 g/L, and 10 g/L for lactose and 20 µM, 100 µM, 500 µM, and 1000 µM for IPTG. The results presented that the concentration of IPTG that obtained the higher antigen levels was that of 100 µM (0.087 g/L), a 10-fold lower concentration than was being previously used in this type of system and for lactose, it was 1 g/L (0.016 g/L). Thus, the induction with 100 µM allowed obtaining the antigen with a concentration 5.6 times higher than the lactose induction maximum concentration.


Subject(s)
Antigens, Protozoan , Escherichia coli/metabolism , Gene Expression , Leishmania infantum , Antigens, Protozoan/biosynthesis , Antigens, Protozoan/genetics , Antigens, Protozoan/immunology , Antigens, Protozoan/isolation & purification , Escherichia coli/genetics , Humans , Leishmania infantum/genetics , Leishmania infantum/immunology , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/isolation & purification
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