| 重组人血管抑素的表达、复性优化及活性研究 |
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| 引用本文： | 陶永辉,赵砚婷,张莲芬,蔡刚明,金坚.重组人血管抑素的表达、复性优化及活性研究[J].中国生化药物杂志,2006,27(1):1-5. |
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| 作者姓名： | 陶永辉 赵砚婷 张莲芬 蔡刚明 金坚 |
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| 作者单位： | 江南大学,生物工程学院,工业生物技术教育部重点实验室,江苏,无锡,214036 |
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| 摘 要： | 目的探索一种实效、经济的血管抑素制备工艺。方法对基因工程菌(E.coli,M15/pQE/AS)的IPTG诱导条件及包涵体复性纯化条件进行优化,产物以SDS-PAGE鉴定纯度,在人微血管内皮细胞模型及鸡胚尿囊膜模型上鉴定其生物学活性。结果经优化后重组人血管抑素的表达量约达菌体蛋白的30%,纯化的重组人血管抑素可显著抑制人微血管内皮细胞的增殖,最高抑制可达47%,并可显著抑制鸡胚尿囊膜模型上血管的生成。结论该表达、复性工艺简单,高效可行。
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| 关 键 词： | 血管抑素 包涵体 人微血管内皮细胞 鸡胚尿囊膜 |
| 文章编号： | 1005-1678（2006）01-0001-05 |
| 收稿时间： | 2005-05-23 |
| 修稿时间： | 2005-07-29 |
Study on the expression,refolding and activity of recombinant human angiostatin |
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| TAO Yong-hui,ZHAO Yan-ting,ZHANG Liang-feng,CAI Gang-ming,JIN Jian.Study on the expression,refolding and activity of recombinant human angiostatin[J].Chinese Journal of Biochemical Pharmaceutics,2006,27(1):1-5. |
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| Authors: | TAO Yong-hui ZHAO Yan-ting ZHANG Liang-feng CAI Gang-ming JIN Jian |
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| Institution: | The Key Lab of Industrial Bioteehnology, Ministry of Education, Southern Yangtze University, Wuxi 214036, China |
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| Abstract: | PurposeTo develop a practicable technology of preparation for recombinant human angiostatin(rhAS).MethodsThe conditions of IPTG inducing to recombinant E.coli(M15/pQE/AS),purifying and refolding to inclusion body were optimized.The purity of rhAS was identified by SDS-PAGE,and the activity was performed on the model of human micro-vascular endothelial cell line(HMEC-1) and chick chorioallantoic membrane.ResultsThe expression of rhAS may achieve 30% of total protein of recombinant E.coli.The best inhibited rate of purified rhAS for HMEC-1 cell line was 47%,and significant inhibition for angiogenesis on chick chorioallantoic membrane was observed.ConclusionIt is a practicable technology of preparation for recombinant human angiostatin(rhAS). |
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| Keywords: | angiostatin inclusion body HMEC-1 chick chorioallantoic membrane |
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