Cloning, Expression, and Refolding of PPE17 Protein of Mycobacterium Tuberculosis as a Promising Vaccine Candidate.
Gene expression
Mycobacterium tuberculosis
Protein refolding
Rv1168c protein
Journal
Iranian journal of medical sciences
ISSN: 0253-0716
Titre abrégé: Iran J Med Sci
Pays: Iran
ID NLM: 8104374
Informations de publication
Date de publication:
Jan 2019
Jan 2019
Historique:
entrez:
23
1
2019
pubmed:
23
1
2019
medline:
23
1
2019
Statut:
ppublish
Résumé
Tuberculosis as a global health problem requires special attention in the contexts of prevention and control. Subunit vaccines are promising strategies to protect burdens of tuberculosis via increasing the BCG protection. The present study aimed to design a vaccine study by means of high-throughput expression and correct refolding of recombinant protein PPE17. We aimed to clone, express, and refold PPE17 protein of The expressed recombinant protein was entirely found in insoluble form (inclusion bodies). The insoluble protein was denatured in 6M guanidine-HCl and refolded by descending denaturant concentration dialysis. Moreover, the recombinant protein was purified by Ni-NTA column chromatography. The changing temperature had no effects on solubilizing protein and the maximum expression was achieved at 0.5 mM concentration of isopropyl-D-thiogalactopyranoside (IPTG) induction. Bacterial expression system is a timesaving tool and refolding by descending denaturant concentration dialysis is a rapid and reliable method.
Sections du résumé
BACKGROUND
BACKGROUND
Tuberculosis as a global health problem requires special attention in the contexts of prevention and control. Subunit vaccines are promising strategies to protect burdens of tuberculosis via increasing the BCG protection. The present study aimed to design a vaccine study by means of high-throughput expression and correct refolding of recombinant protein PPE17.
METHODS
METHODS
We aimed to clone, express, and refold PPE17 protein of
RESULTS
RESULTS
The expressed recombinant protein was entirely found in insoluble form (inclusion bodies). The insoluble protein was denatured in 6M guanidine-HCl and refolded by descending denaturant concentration dialysis. Moreover, the recombinant protein was purified by Ni-NTA column chromatography. The changing temperature had no effects on solubilizing protein and the maximum expression was achieved at 0.5 mM concentration of isopropyl-D-thiogalactopyranoside (IPTG) induction.
CONCLUSION
CONCLUSIONS
Bacterial expression system is a timesaving tool and refolding by descending denaturant concentration dialysis is a rapid and reliable method.
Types de publication
Journal Article
Langues
eng
Pagination
53-59Références
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