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首页> 外文期刊>Protein Expression and Purification >Producing human mechano growth factor (MGF) in Escherichia coli
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Producing human mechano growth factor (MGF) in Escherichia coli

机译：在大肠杆菌中产生人类机械生长因子（MGF）

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MGF is a product of a unique muscle-specific splice variant of IGF1 gene (insulin-like growth factor). Its peculiar feature is a specific E-peptide, a 16 a.a. strand at the C-terminus. MGF increases cellular proliferation and inhibits terminal differentiation of myoblasts necessary for the secondary myotube formation. Previous analysis of physiological effects of MGF was performed using indirect methods such as RT-PCR based examination of the transcript contents in normal tissues, adenovirus-mediated DNA delivery and synthetic E-domain administration. Here, we describe isolation and purification of recombinant MGF thus allowing for the first time the possibility of direct examining MGF effects. The recombinant MGF of directly examining-was expressed in Escherichia coli as inclusion bodies (about 100-200 mg/l), purified and refolded. Biological activity of refolded MGF was analyzed in vitro in proliferation assays with normal human myoblasts. As a result of our work, it has become possible to generate a standard MGF control with characterized activity and a ready-to use MGF test-system neither of which have been previously described. Our data open opportunities for the future works on MGF characterization and to the development of a powerful and highly specific therapeutic agent potentially applicable for muscle growth up-regulation, post-trauma muscle repair, age and hereditary myodystrophy mitigation and in sport medicine. (c) 2007 Elsevier Inc. All rights reserved.
机译：MGF是IGF1基因（胰岛素样生长因子）独特的肌肉特异性剪接变体的产物。它的独特特征是特异性的E肽，a.a.a。16。在C末端MGF增加细胞增殖并抑制继发性肌管形成所必需的成肌细胞的终末分化。使用间接方法，例如基于RT-PCR的正常组织中转录物含量检查，腺病毒介导的DNA传递和合成E结构域给药，对MGF的生理效应进行了先前的分析。在这里，我们描述了重组MGF的分离和纯化，因此首次使直接检查MGF效果成为可能。直接检查的重组MGF在大肠杆菌中表达为包涵体（约100-200 mg / l），纯化并重折叠。在正常人成肌细胞的增殖试验中，体外分析了重折叠的MGF的生物学活性。作为我们工作的结果，已经可以生成具有特征性活动的标准MGF控件和现成的MGF测试系统，而这两种功能均未在前面进行描述。我们的数据为MGF的鉴定和功能强大且高度特异性的治疗剂的未来开发提供了机会，这些治疗剂可能适用于肌肉生长上调，创伤后肌肉修复，年龄和遗传性肌营养不良的缓解以及运动医学。 （c）2007 Elsevier Inc.保留所有权利。

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