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目的 为了获得大量重组人血管内皮细胞生长因子（ Ｈｕｍａｎ Ｖａｓｃｕｌａｒ ＥｎｄｏｔｈｅｌｉａｌＧｒｏｗｔｈ Ｆａｃｔｏｒ， ｈＶＥＧＦ），以研究其在心血管和药学领域具有的潜在药用价值。方法 用ＰＣＲ方法扩增目的基因ｈＶＥＧＦ１６５，连接到 ｐＥＴ－３ａ载体上，并转化到大肠杆菌ＢＬ２１（ＤＥ３）中，建立可高效表达的ｐＥＴ－３ａ／ｈＥＶＧＦ１６５重组质粒，经ＩＰＴＧ诱导表达， Ｗｅｓｔｅｒｎ ｂｌｏｔ检测表达产物的抗原性，通过 Ｍｏｎｏ Ｑ阴离子层析和ＦＰＬＧ Ｓｕｐｅｒｏｓｅ１２分子筛柱层析，进行初步纯化，用 ＭＴＴ法检测其生物学活性。结果 ｐＥＴ－３ａ／ｈＥＶＧＦ表达产物经纯化后，其纯度可达到９０％以上，表达的ｒｈＥＶＧＦ１６５能 呈剂量依赖性地促进人静脉内皮细胞（ＨＵＶＥＣ）的增殖。结论组建了天然形式ｈＥＶＧＦ１６５的大肠杆菌菌株，并摸索 出一套纯化工艺，为大规模生产重组ｈＥＶＧＦ１６５奠定了基础。
Objective To obtain a large number of human vascular endothelial growth factor (hVEGF) in order to study its potential cardiovascular and pharmacological properties. Methods The target gene hVEGF165 was amplified by PCR, ligated into pET-3a vector, and transformed into E. coli BL21 (DE3). The recombinant plasmid pET-3a / hEVGF165 was constructed and expressed by IPTG. The antigenicity of the expressed product was determined by Mono Q anion chromatography and FPLG Superose 12 molecular sieve column chromatography. The biological activity of the product was tested by MTT assay. Results The purity of pET-3a / hEVGF was over 90% after purification. The expressed rhEVGF165 could promote the proliferation of human vein endothelial cells (HUVECs) in a dose-dependent manner. Conclusion The E. coli strain of natural form hEVGF165 was established and a purification process was developed, which laid the foundation for large-scale production of recombinant hEVGF165.