Estratégias de indução do antígeno 503 recombinante de Leishmania i. chagasi expresso em Escherichia coli M15
With nearly 900,000 to 1.3 million new cases annually, 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 disease in humans so far, and its treatment is expensive and...
में बचाया:
| मुख्य लेखक: | |
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| अन्य लेखक: | |
| स्वरूप: | Dissertação |
| भाषा: | por |
| प्रकाशित: |
Brasil
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| विषय: | |
| ऑनलाइन पहुंच: | https://repositorio.ufrn.br/jspui/handle/123456789/25672 |
| टैग : |
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| सारांश: | With nearly 900,000 to 1.3 million new cases annually, 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 disease in humans so far, and its treatment is expensive
and aggressive to human health. Vaccination remains the most practical and realistic way of
fight against the disease, thus, the production of recombinant antigens as a pathway towards a
vaccine acquisition is necessary. In addition, the optimization of the strategy of the recombinant
antigen’s production is of interest so it can make the process viable. Therefore, the present study
aims to optimize the induction parameters of the 503 Leishmania i. chagasi antigen expressed
in recombinant Escherichia coli. Hence, the induction at different cell densities (optical density
at 600 nm of 0.5 and 1.0) was evaluated in order to analyse the influence of the induction time
on the yield of the protein of interest. In this segment, lactose and isopropyl-β-Dthiogalactopyranoside
(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 to obtain the antigen with a concentration 5.6 times higher than the lactose induction
maximum concentration. |
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