Bibliographic citations
Rios, A., (2021). Evaluación y comparación de la concentración y solubilidad de la proteína ClpC1 de Mycobacterium tuberculosis producida de manera recombinante mediante el uso de los plásmidos pET28a(+) y pMAL-cHT en tres cepas distintas de Escherichia coli [Universidad Peruana Cayetano Heredia]. https://hdl.handle.net/20.500.12866/9711
Rios, A., Evaluación y comparación de la concentración y solubilidad de la proteína ClpC1 de Mycobacterium tuberculosis producida de manera recombinante mediante el uso de los plásmidos pET28a(+) y pMAL-cHT en tres cepas distintas de Escherichia coli []. PE: Universidad Peruana Cayetano Heredia; 2021. https://hdl.handle.net/20.500.12866/9711
@misc{sunedu/4050894,
title = "Evaluación y comparación de la concentración y solubilidad de la proteína ClpC1 de Mycobacterium tuberculosis producida de manera recombinante mediante el uso de los plásmidos pET28a(+) y pMAL-cHT en tres cepas distintas de Escherichia coli",
author = "Rios Angulo, Angela Anaid",
publisher = "Universidad Peruana Cayetano Heredia",
year = "2021"
}
Clp proteins are a group of caseolytic proteases of the ATP-dependent serine group. Its main function is the quality control of proteins. In Mycobacterium tuberculosis, ClpC1 participates in the mechanism of action of antituberculous drugs such as pyrazinamide (PZA), which is why studies on its function and structure are important. Also, ClpC1 monomer from Bacillus subtilis has been completely crystallized so it is used as a template to model ClpC1 from other bacteria. ClpC1 is a cytosolic protein so its expression in soluble form has been confirmed in previous studies. The expression system in Escherichia coli using the plasmid pET28a (+) as an expression vector has been used to produce ClpC1; however, there is no information about the percentage of solubility and concentration of protein produced. On the other hand, the plasmid pMAL-cHT is widely used to increase the solubility of recombinant proteins in E. coli. pMAL-cHT adds the malE gene, which encodes the maltose-binding protein (MBP), to the protein to be expressed and, by mechanisms not yet confirmed, increases the solubility and quantity of recombinant proteins. In this study, the concentration and percentage of solubility of M. tuberculosis ClpC1 expressed in different expression systems were estimated using vectors that included or did not include the MBP fusion protein, different concentrations of inducer and induction times. The best conditions to obtain a higher concentration and percentage of solubility of ClpC1 were with the strain BL21 (DE3) pLysS, the plasmid pMAL-cHT-clpC1 induced with 0.5 mM of IPTG for 10 hours at 16 ° C. In this system, 45.54 mg / ml (95% CI 44.68 - 46.40) of ClpC1 was obtained with a solubility percentage of 86.93% (95% CI 80.41 - 93.45). In the same way, the bioinformatic model of ClpC1 with MBP presented greater global solubility compared to ClpC1.
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