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为了克隆表达鸡的基质金属蛋白酶 2 (MMP 2 )的C端片段PEX ,并探讨其对血管发生的抑制作用 ,利用RT PCR从鸡胚成纤维细胞克隆MMP 2C端片段PEX ,构建原核表达载体pCal n PEX ;转化大肠杆菌BL2 1(DE3) pLys,异丙基β D 硫代半乳糖苷 (IPTG)诱导产生PEX融合蛋白 ,包涵体蛋白用盐酸胍法变性、复性 ;生长曲线观察PEX融合蛋白对人脐静脉血管内皮细胞增殖的影响 ;鸡胚绒毛尿囊膜血管发生实验研究其对血管发生的抑制作用 .结果表明融合蛋白CBP/PEX具有抑制人脐静脉血管内皮细胞的生长和鸡胚绒毛尿囊膜血管发生的作用 .提示PEX是有待进一步开发的潜在抑制血管发生的药物 .
In order to clone the C-terminal fragment of matrix metalloproteinase 2 (MMP 2) and investigate its inhibitory effect on angiogenesis, RT-PCR was used to clone the MMP 2C end fragment from chick embryo fibroblasts to construct prokaryotic expression vector pCal nPEX. The PEX fusion protein was induced by isopropyl β D thiogalactopyranoside (IPTG) after transformed into E. coli BL21 (DE3) pLys. Inclusion body protein was denatured and refolded with Guanidine Hydrochloride. The growth curve was used to observe the expression of PEX fusion protein On the proliferation of human umbilical vein endothelial cells; chick chorioallantoic membrane angiogenesis experimental study of its angiogenesis inhibition.The results show that the fusion protein CBP / PEX can inhibit the growth of human umbilical vein endothelial cells and chick embryo villi The role of allantoic membrane angiogenesis suggests that PEX is a potential drug for angiogenesis to be further developed.