Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Expression and purification of recombinant TAT-BoNT/A((1-448)) under denaturing and native conditions

(2016) Expression and purification of recombinant TAT-BoNT/A((1-448)) under denaturing and native conditions. Bioengineered. pp. 478-483. ISSN 2165-5979

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Expression and purification of recombinant TAT-BoNTA((1-448)) under denaturing and native conditions.pdf

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Abstract

Botulinum toxin type A can temporarily inhibit muscle contraction. Currently, physicians administer this toxin as a bio-drug in treatment of some muscle contraction disorders. TAT-BoNT/A((1-448)) is a functional recombinant protein derived from botulinum toxin light chain. Unlike the full length botulinum toxin, TAT-BoNT/A((1-448)) is a self-permeable molecule which can pass through bio-surfaces so can be used as a topical therapeutic agent without injection. To maintain the functionality of TAT-BoNT/A((1-448)), it is necessary to restore its normal folding upon expression and purification. In this study, we have investigated and optimized expression conditions for this novel recombinant protein. Under denaturing condition (1mM IPTG, at 37 degrees C), the chimeric protein was produced as inclusion body and required to be purified using denaturing agents (e.g. urea). Yet, lower incubation temperature (18 degrees C) and less IPTG concentration (0.5mM) induce a protein under native condition. In such condition, about 60 of the chimeric protein was expressed in soluble form.

Item Type: Article
Keywords: cell-penetrating peptide inclusion body light chain of botulinum toxin type A protein expression TAT-BoNT A((1-448)) clostridium-botulinum structural-analysis neurotoxins toxin Biotechnology & Applied Microbiology
Divisions:
Page Range: pp. 478-483
Journal or Publication Title: Bioengineered
Journal Index: ISI
Volume: 7
Number: 6
Identification Number: https://doi.org/10.1080/21655979.2016.1201252
ISSN: 2165-5979
Depositing User: مهندس مهدی شریفی
URI: http://eprints.bmsu.ac.ir/id/eprint/5230

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