Banca de QUALIFICAÇÃO: VITOR TROCCOLI RIBEIRO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : VITOR TROCCOLI RIBEIRO
DATA : 21/12/2017
HORA: 14:00
LOCAL: SALA DO CATRE
TÍTULO:

"Evaluation of the pQE-30 plasmid stability and expression of 503 antigen of Leishmania i. chagasi in Escherichia coli M15 at different concentrations of IPTG and culture temperatures."

PALAVRAS-CHAVES:

Leishmania i. chagasi, induction, temperature, 503 antigen, plasmidial stability.

PÁGINAS: 83
GRANDE ÁREA: Engenharias
ÁREA: Engenharia Química
RESUMO:

Visceral leishmaniasis is an infectious-parasitic disease caused by protozoa of the genus Leishmania, which can be lethal when there is no adequate treatment. It is estimated that there are 12 million people infected by the disease, but unfortunately there are no drugs capable of preventing the disease. There are drugs for the treatment of the disease, but these have a high cost and high toxicity. So it is necessary to look for more effective alternatives, less aggressive and low cost. It is attempted to achieve this goal through the production of antigens from Leishmania i. chagasi, using Escherichia coli as host for genetic recombination, being a well characterized prokaryote and of known metabolism. In this context, the present work aims to evaluate the influence of the concentration of IPTG during the induction and the influence of the culture temperature on 503 antigen expression and on the stability of plasmid pQE-30. All assays were performed in a rotary incubator and made in duplicate. First, IPTG induced cultures were carried out at concentrations of: 0.01 mM; 0.1 mM; 0.5 mM; 1.0 mM and 1.5 mM at 37 °C. The concentration of the inducer was not a factor that affected the stability of the plasmid and when induced with 1.0 mM, the 503 antigen expression was maximal 0.101 ± 0.004 g / L. It was possible to verify that the lowest concentration of IPTG used was enough to achieve expression of the recombinant protein. For the next assays, the concentration of 1.0 mM IPTG was fixed and the culture temperatures used were 27, 32 and 42 °C. These assays were compared with the previous assay performed at 37 ° C. Temperature was a factor that influenced the stability of the plasmid, the lower the culture temperature, the greater the stability. Even so, the temperature of 37 ° C was still the one that obtained greater expression of 503 antigen.

 

MEMBROS DA BANCA:
Externo ao Programa - 2378605 - CRISTIANE FERNANDES DE ASSIS
Presidente - 1346198 - EVERALDO SILVINO DOS SANTOS
Externo ao Programa - 3652554 - FRANCISCO CANINDE DE SOUSA JUNIOR