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目的 在大肠杆菌中表达赖型钩端螺旋体毒力相关蛋白InvA ,并观察该蛋白对ANA 1细胞的增殖作用。方法 将invA基因克隆于pET32a(+)载体 ,在大肠杆菌BL2 1(DE3)中表达带组氨酸标签的Trx InvA融合蛋白 ,经亲和层析获得纯化Trx InvA蛋白。以不同浓度的Trx InvA融合蛋白作用于ANA 1细胞 ,以四氮唑复合物 [2 ,3 bis(2 methoxy 4 nitro sulfophenyl) 5 〔(Phenylamino)carbonyl〕 2H tetrazoiumhydrixide,XTT]法检测细胞增殖的变化 ,分析InvA蛋白的细胞功能。结果 成功构建pETIN VA原核表达载体 ,在大肠杆菌BL2 1(DE3)中表达了相对分子质量 (Mr)约为 35× 10 3的Trx InvA融合蛋白 ,该蛋白能促进ANA 1细胞的增殖。结论 表达并纯化的Trx InvA融合蛋白具有促进细胞增殖的作用
Objective To express InvA of Leptospira interrogans virus in Escherichia coli and observe the proliferation of ANA 1 cells. Methods The invA gene was cloned into the pET32a (+) vector. Histidine tagged Trx InvA fusion protein was expressed in Escherichia coli BL21 (DE3). The purified Trx InvA protein was obtained by affinity chromatography. ANA1 cells were treated with different concentrations of Trx InvA fusion protein and the changes of cell proliferation were detected by tetrazolium complex [2,3 bis (2 methoxy 4 nitro sulfophenyl) 5 [(Phenylamino) carbonyl] 2H tetrazoiumhydrixide , Analyze the cell function of InvA protein. Results The prokaryotic expression vector pETIN VA was successfully constructed. The Trx InvA fusion protein with molecular weight (Mr) of 35 × 10 3 was expressed in Escherichia coli BL21 (DE3), which can promote the proliferation of ANA1 cells. Conclusion The expressed and purified Trx InvA fusion protein has the effect of promoting cell proliferation