骨形态发生蛋白-2的高效表达纯化和复性-食品科学.PDFVIP

190 2013, Vol.34, No.15 食品科学 ※生物工程 骨形态发生蛋白-2的高效表达、纯化和复性 1 2 1 1 杨春瑜 ，杨春莉，刘晓飞，赫 美 (1.黑龙江省普通高校食品科学与工程省重点实验室，哈尔滨商业大学食品工程学院，黑龙江 哈尔滨 150076 ； 2.哈尔滨商业大学轻工学院，黑龙江 哈尔滨 150028) 摘  要：从含BMP-2cDNA质粒的E.coli 中获得骨形态发生蛋白-2(BMP-2) 的目的基因，构建质粒pET28a-BMP-2并转 入BL 2 1(DE3) E.coli 中，经酶切鉴定，证明BMP-2在BL 2 1(DE3) E.coli 中成功诱导表达。摸索高效表达条件后，对 BMP-2进行纯化和复性，得到具有较高生物活性的BMP-2 。采用低成本方法解决骨修复材料和BMP-2缓释研究中大 量价格昂贵的BMP-2需求问题。 关键词：BMP-2 ；诱导表达；pET-28 a(+) ；纯化；复性 Optimal Expression, Purification and Renaturation of BMP-2 in E.coli BL 21(DE3) 1 2 1 1 YANG Chun-yu ，YANG Chun-li ，LIU Xiao-fei ，HE Mei (1. Key Laboratory of Food Science and Engineering of Heilongj iang Province, College of Food Engineering, Harbin University of Commerce, Harbin 150076, China ；2. College of Light Industry, Harbin University of Commerce, Harbin 150028, China) Abstract ：The plasmid pET28a-BMP-2 based on the target gene of bone morphogenetic protein-2 (BMP-2) from E.coli harboring plasmid BMP-2cDNA was transformed into E.coli BL 21(DE3). The successful expression of BMP-2 in E.coli BL 21(DE3) was identified by enzymatic digestion. Under optimized expression conditions, BMP-2 was purified and renaturated to obtain enhanced bioactivity. This study provides a cost-effective way of producing BMP-2, which is expensive and needed in large quantities in bone repair materials and investigation of its sustained release capacity. Key words ：bone morphogenetic protein -2 ；inducement expression ；pet-28 a(+) ；purification ；renaturation 中图分类号：TS20 1.1 文献标志码：A 文章编号：1002-6630(2013)15-0190-04 doi:10.7506/spkx 1002-6630-20 1