重组人血管抑素的表达、复性优化及活性研究  

Study on the expression,refolding and activity of recombinant human angiostatin

作　　者：陶永辉[1] 赵砚婷[1] 张莲芬[1] 蔡刚明[1] 金坚[1] 

机构地区：[1]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

出　　处：《中国生化药物杂志》2006年第1期1-5,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：江苏省自然科学基金(No.BK2002071)

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。Purpose To develop a practicable technology of preparation for recombinant human angiostatin （rhAS）. Methods The conditions of IPTG inducing to recombinant E. coli （ M 15/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 chofioallantoic membrane. Results The expression of rhAS may achieve 30% of total protein of recombinant E. coll. 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. Conclusion It is a practicable technology of preparation for re combinant human angiostatin（rhAS）.

关 键 词：血管抑素 包涵体 人微血管内皮细胞 鸡胚尿囊膜 

分 类 号：Q516[生物学—生物化学] Q786