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CiteScore (2016): 2.10

SNIP (2016): 1.075

SJR (2016): 0.61

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Adv Pharm Bull. 2016;6(2):187-194.
doi: 10.15171/apb.2016.026
PMID: 27478780
PMCID: PMC4961976
Scopus id: 84980383507
  Abstract View: 431
  PDF Download: 245

Research Article

Cloning and Expression of Recombinant Human Endostatin in Periplasm of Escherichia coli Expression System

Abbas Mohajeri 1, Yones Pilehvar-Soltanahmadi 1,2, Mohammad Pourhassan-Moghaddam 2, Jalal Abdolalizadeh 3, Pouran Karimi 4, Nosratollah Zarghami 1,2 *

1 Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Department of Medical Biotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Neurosciences Research Center (NSRC), Tabriz University of Medical Sciences, Tabriz, Iran.

Abstract

Purpose: Recombinant human endostatin (rhEs) is an angiogenesis inhibitor which is used as a specific drug in the treatment of non-small-cell lung cancer. In the current research, we developed an efficient method for expressing soluble form of the rhEs protein in the periplasmic space of Escherichia coli via fusing with pelB signal peptide.

Methods: The human endostatin (hEs) gene was amplified using synthetic (hEs) gene as a template; then, cloned and expressed under T7 lac promoter. IPTG was used as an inducer for rhEs expression. Next, the osmotic shock was used to extraction of protein from the periplasmic space. The presence of rhEs in the periplasmic space was approved by SDS-PAGE and Western blotting.

Results: The results show the applicability of pelB fusion protein system usage for secreting rhEs in the periplasm of E. coli in the laboratory scale. The rhEs represents approximately 35 % (0.83mg/l) of the total cell protein.

Conclusion: The present study apparently is the first report of codon-optimized rhEs expression as a fusion with pelB signal peptide. The results presented the successful secretion of soluble rhEs to the periplasmic space.

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