人血管内皮生长因子VEGF183(132-158)肽段含有细胞外基质结合序列与纤溶酶、基质金属蛋白酶以及uPA酶裂解位点,研究该肽段作为一种蛋白药物的缓释载体,在体内既可和细胞外基质结合,又可在体内的相应酶作用下,缓慢释放。该研究以绿色荧光蛋白GFP为指示分子,进一步采用加端PCR方法将VEGF183(132-158)肽段连到GFP的C端,原核表达并纯化该融合蛋白,通过与琼脂糖肝素柱结合实验与冷冻切片的荧光显微镜观察,发现该肽段赋予了GFP蛋白结合琼脂糖肝素柱与细胞外基质的功能,GFP融合蛋白的获得为下一步的体内释放研究打下了基础。 The human vascular endothelial growth factor VEGF183 (132-158) peptide contains extracellular matrix-binding sequences and plasmin, matrix metalloproteinase and uPA cleavage sites. As a sustained release carrier of protein drugs, this peptide has been studied in vivo Can be combined with the extracellular matrix, but also in the body under the action of the corresponding enzyme, the slow release. In this study, green fluorescent protein GFP was used as an indicator molecule. The VEGF183 (132-158) peptide was further linked to the C-terminal of GFP using an additive-end PCR method. The fusion protein was expressed in prokaryotic cells and purified by agarose heparin column Fluorescence microscopy of frozen sections showed that this peptide conferred the function of GFP binding agarose heparin column and extracellular matrix. The gain of GFP fusion protein laid the foundation for the next in vivo release study.