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目的将X连锁凋亡抑制蛋白(XIAP)插入质粒pTAT-HA,构建pTAT-XIAP原核表达载体,以获得高产量、高纯度的pTAT-XIAP融合蛋白。方法将pTAT-XIAP融合蛋白表达载体转入大肠杆菌BL21plysS,异丙基硫代半乳糖苷(IPTG)诱导融合蛋白表达,产物经镍离子亲和层析柱(Ni-NTA)亲和层析纯化,并用Western blot方法鉴定。结果 pTAT-XIAP融合蛋白在大肠杆菌中获得表达,纯化后蛋白呈单一条带,表达产物经Western blot分析证实目的蛋白表达。结论构建重组融合表达载体,在大肠杆菌中成功表达并纯化pTAT-XIAP融合蛋白。
Objective To construct the prokaryotic expression vector pTAT-XIAP by inserting XIAP into plasmid pTAT-HA to obtain high-yield and high-purity pTAT-XIAP fusion protein. Methods The pTAT-XIAP fusion protein expression vector was transformed into E. coli BL21plysS and induced by IPTG. The fusion protein was purified by Ni-NTA affinity chromatography , And identified by Western blot. Results The pTAT-XIAP fusion protein was expressed in E. coli. The purified protein showed a single band. The expressed protein was confirmed by Western blot. Conclusion The recombinant fusion expression vector was constructed and the pTAT-XIAP fusion protein was successfully expressed and purified in E. coli.