人血管生成抑制素基因在大肠杆菌中的融合表达

Human Angiostatin:Its High level Expression in E coli

将人血管生成抑制素基因定向插入表达载体ｐＥＴ－１７ｂ的ＢａｍＨⅠ位点，构建重组质粒ｐＥＴＡ２，转化Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）．在ＩＰＴＧ诱导下，血管生成抑制素基因在重组转化株Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３，ｐＥＴＡ２）中获得高效表达．表达产物以包涵体形式存在，表达量占菌体总蛋白的３７．２％．利用羊抗人纤溶酶原抗血清作为第一抗体。

Angiostatin is a recently discovered angiogenesis inhibitor, capable in inhibitting the proliferation of endothelial cells and the growth of tumor and has the potential for clinical application.Human angiostatin gene was cloned on the BamHI site of pET 17b resulting in the plasmid PETA 2 which was then transformed into E coli BL21(DE3). Angiostatin was efficently expressed in recombinant strain E coli BL21(DE3, pETA2) by IPTG induction. The expression product constitutes about 37 2% of the total soluble protein and exsists in the form of inclusion body. Western blot analysis demonstrates that it reacts positively with goat anti human plasminogen antisera.