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目的:利用原核表达系统获得重组血管生成素 (Ang)并研究其生物学活性。方法:构建高效融合表达载体pGEX 4T Ang,诱导表达后利用亲和层析的方法纯化目的蛋白,经切割后获得非融合的重组人Ang,利用Western印迹检测表达产物的特异性,通过鸡胚尿囊膜血管增生实验检测重组蛋白的生物学功能。结果:表达的目的蛋白约占菌体总蛋白的 20%,亲和层析纯化蛋白的纯度在 90%,能够与Ang抗体产生特异性反应,并具有明显的促进血管新生的活性。结论:原核表达的重组Ang具有良好的生物学活性。
Objective: To obtain recombinant Angiopoietin (Ang) by using prokaryotic expression system and to study its biological activity. Methods: The fusion protein pGEX 4T Ang was constructed. After induced expression, the target protein was purified by affinity chromatography. The fusion protein was purified by Western blotting. The specificity of the expressed product was detected by Western blot. Capillary Vascular Hyperplasia Experiment to Detect the Biological Function of Recombinant Protein. Results: The expressed protein accounted for about 20% of the total bacterial proteins, and the purity of the purified protein was 90%. It could react specifically with Ang antibody and had obvious angiogenic activity. Conclusion: Recombinant prokaryotic expression of Ang has good biological activity.