摘要
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)
Objective To construct an engineering bacteria which can express human endostatin protein with biological activity. Methods Human endostatin gene fragment was acquired from human fetal liver total RNA by means of RT PCR. The gene was inserted into prokaryotic expressing vector pBV220 and transformed into E.coli DH5α. Endostatin was expressed in the E.coli and the inclusion body was isolated, solubilized and then refolded. Results SDS PAGE analysis showed that expressed product was about M r 20×10 3 and took up about 30% in bacterial total protein. Conclusion The recombinant human endostatin protein can inhibit proliferation of bovine capillary endothelial cells dependent on bFGF.
出处
《中华微生物学和免疫学杂志》
CSCD
北大核心
2000年第5期402-404,共3页
Chinese Journal of Microbiology and Immunology
