人血管抑制因子vasostatin片段在大肠杆菌中的融合表达、纯化及活性检测

发布时间：2018-03-16 22:51

本文选题：vasostatin片段　切入点：GST融合蛋白　出处：《感染.炎症.修复》2005年01期 　论文类型：期刊论文

【摘要】：目的：克隆表达人vasostatin片段,纯化目的蛋白并检查活性,以研究人血管抑制因子vasostatin片段的活性区域。方法：重组构建人vasostatin片段和谷胱甘肽S转移酶(GST)融合表达质粒,转化大肠杆菌BL21 (DE3),IPTG诱导表达。通过GST亲和凝胶纯化目的蛋白,用凝血酶(Thrombin)酶切纯化产物获得单体va- sostatin片段,并采用MTT法检测其抑制内皮细胞增殖活性。结果：表达产物主要以包涵体形式存在,改变诱导条件可增加可溶性表达量,包涵体形式的表达量约占菌体蛋白总量的55%。活性试验表明,各GST-vasostatin片段和vasoatatin片段具有程
[Abstract]:Objective: to clone and express human vasostatin fragment, purify the target protein and examine the activity of vasostatin fragment. Methods: the fusion expression plasmid of human vasostatin fragment and glutathione S-transferase (GST) was constructed. The target protein was purified by GST affinity gel and purified by thrombin enzyme digestion to obtain monomer va- sostatin fragment. MTT assay was used to detect the inhibition of endothelial cell proliferation. Results: the expression product mainly existed in the form of inclusion body, and the soluble expression could be increased by changing the induction condition. The expression of inclusion body accounted for about 55% of the total bacterial protein. The activity test showed that each GST-vasostatin fragment and vasoatatin fragment had a sequence.
【作者单位】：暨南大学医药生物技术研究开发中心暨南大学医药生物技术研究开发中心暨南大学医药生物技术研究开发中心解放军全军创伤修复重点实验室暨南大学医药生物技术研究开发中心
【分类号】：R346

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