Mohajeri Abbas, Pilehvar-Soltanahmadi Yones, Pourhassan-Moghaddam Mohammad, Abdolalizadeh Jalal, Karimi Pouran, Zarghami Nosratollah
Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.; Department of Medical Biotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2016 Jun;6(2):187-94. doi: 10.15171/apb.2016.026. Epub 2016 Jun 30.
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.
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.
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.
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.
重组人内皮抑素（rhEs）是一种血管生成抑制剂，用作治疗非小细胞肺癌的特效药物。在本研究中，我们开发了一种通过与pelB信号肽融合，在大肠杆菌周质空间高效表达可溶性rhEs蛋白的方法。
以合成的人内皮抑素（hEs）基因作为模板扩增hEs基因；然后，在T7 lac启动子下进行克隆和表达。IPTG用作rhEs表达的诱导剂。接下来，采用渗透压休克法从周质空间提取蛋白质。通过SDS-PAGE和蛋白质免疫印迹法验证周质空间中rhEs的存在。
结果表明，在实验室规模下，pelB融合蛋白系统可用于在大肠杆菌周质中分泌rhEs。rhEs约占总细胞蛋白的35%（0.83mg/l）。
本研究显然是关于密码子优化的rhEs与pelB信号肽融合表达的首次报道。结果表明可溶性rhEs成功分泌到周质空间。
