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目的 获得具有抑制血管内皮细胞生长活性的重组人内皮抑素 (recombinanthumanen dostatin)。方法 从肝细胞中分离总RNA ,经反转录多聚酶链反应 (RT PCR)得到endostatin全基因。用原核细胞表达载体构建了pBV2 2 0 /endostatin重组质粒 ,经DNA测序确认后 ,将其转化大肠杆菌DH5α进行表达、初步纯化及活性测定。结果 经SDS PAGE分析 ,表达产物相对分子质量 (Mr)约 2 0× 10 3,薄层扫描显示表达产物可达细菌总蛋白的 30 %。结论 经生物学活性测定 ,表达蛋白能够抑制碱性成纤维细胞生长因子 (bFGF)对牛毛细血管内皮细胞 (BCEC)的增殖作用
Objective To obtain recombinant human interleostatin with inhibitory effect on the growth of vascular endothelial cells. Methods Total RNA was isolated from hepatocytes and all the endostatin genes were obtained by reverse transcription polymerase chain reaction (RT PCR). The recombinant plasmid pBV220 / endostatin was constructed by using prokaryotic expression vector. After confirmed by DNA sequencing, it was transformed into E. coli DH5α for expression, preliminary purification and activity determination. Results The relative molecular mass (Mr) of the expressed product was about 20 × 10 3 by SDS PAGE. The thin layer scanning showed that the expressed product reached 30% of the total bacterial protein. Conclusion The biological activity of the expressed protein can inhibit the proliferation of bovine capillary endothelial cells (BCEC) induced by basic fibroblast growth factor (bFGF)