中文摘要：

目的首次报道利用东亚钳蝎镇痛抗肿瘤缬精甘肽-BmKAGAP产生两种不同的融合蛋白，目的是获得镇痛和抗肿瘤的双功能活性。方法融合蛋白是以嵌合毒素的形式进行构建，由三部分组成，包括促性腺激素释放激素（1uteinizing hormone-releasing hormoneL，HRH），两种不同类型的柔性连接物，以及BmKAGAP。克隆编码两种融合蛋白的基因并在大肠杆菌中实现可溶性表达。通过金属离子螯合亲和层析和阳离子交换层析方法对融合蛋白分离纯化，采用小鼠醋酸扭体法和MTT法测定融合蛋白的镇痛活性与细胞抑制活性。结果融合蛋白在小鼠模型中体现出镇痛活性（rLHRH-Tag（His）-AGAP〉rTagfHis）-LHRH-L8-AGAP〉rBmKAGAP），同时对肝癌细胞株Hep3B具有细胞毒活性（rLHRH-Tag（His）-AGAP〉rBmKAGAP〉rTag（His）∥LHRH-L8-AGAP）。结论得到优化的双功能融合蛋白候选者为嵌合毒素rLHRH-Tag（His）-AGAP。

英文摘要：

Objective To obtain two different fusion proteins with an antitumor-analgesic peptide from Chi- nese scorpion Buthus martensii Karsch （BmKAGAP）, aiming to acquire bifunctional analgesic and antitu- mor activity. Methods The fusion proteins were constructed in the form of chimeric toxins. The fusion pro- teins consisted of luteinizing hormone-releasing hormone （LHRH）, two different types of flexible linkers, and BmKAGAP. The genes coding two fusion proteins were cloned and were expressed in E. coli BL21 （ADE3）, and purified by metal chelating affinity chromatography and ion exchange chromatography. The analgesic and cytotoxic activity were detected by the mouse twisting action model and MTT assay, respec- tively. Results The fusion proteins showed analgesic activity （rLHRH-Tag（His） -AGAP 〉 rTag（His） -LHRH- L8-AGAP 〉 rBmKAGAP）, they also have cytotoxic activity to hepatocellular carcinoma cell line Hep3B （rLHRH-Tag（His）-AGAP 〉 rBmKAGAP 〉 rTag（His）-LHRH-L8-AGAP）. Conclusions An optimized bifunc- tional fusion protein candidate is obtained,namely the chimeric toxin of rLHRH-Tag（His）-AGAP.