三叶半夏凝集素C端结构域的可溶表达及其活性研究

利用pET-32-SUMO表达载体在大肠杆菌中可溶表达三叶半夏凝集素C端结构域（P-D2）,并研究其凝血活性、糖结合特异性以及肿瘤抑制活性。实验结果表明,P-D2蛋白的产量为5mg/L。该蛋白具有凝集小鼠红细胞的活性,可以被D-甘露聚糖抑制,但不能被D-乳糖、D-葡萄糖、D-麦芽糖抑制。P-D2蛋白的体外抗肿瘤活性实验表明其对肝癌细胞Bel-7404以及结肠癌细胞SW-620都有良好的抑制作用。凋亡现象观察实验发现P-D2蛋白可以诱导癌细胞Bel-7404的凋亡。这是首次在大肠杆菌表达系统中表达了可溶的半夏凝集素C端结构域,为其他凝集素的可溶表达提供了新的选择。

Study on Soluble Expression and Bioactivity of C-Terminus Domain of Pinellia Ternata Agglutinin

pET-32-SUMO expression vector was used for soluble expression of C-terminus domain of pinellia ternata agglutinin（P-D2） and study on hemagglutination activity,saccharides binding specificity and tumor-inhibition activity.The results show that,the output of P-D2 protein is 5mg/L.This protein has the activity to agglutinate erythrocyte of the mice and can be inhibited by D-mannan but not by D-lactose,D-glucose and D-maltose.The experiment of antitumor activity in vitro of P-D2 protein indicates it has good inhibiting effect on hepatoma carcinoma cell Bel-7404 and colon cancer cell SW-620.It is found through observation of apoptosis phenomenon that P-D2 can induce apoptosis of Bel-7404.Soluble C-terminus domain of pinellia ternata agglutinin is expressed in escherichia coli expression system the first time.This provides new choice for soluble expression of other agglutinin.