【摘要】 目的体外制备刚地弓形虫醛缩酶(aldolase)蛋白并纯化。方法以刚地弓形虫cDNA链为模板,聚合酶链反应扩增aldolase基因,克隆至质粒pGEX-4T-1上,转化至大肠杆菌BL21;异丙基-β-D-硫代半乳糖苷诱导表达,亲和层析纯化表达产物。结果获得了aldolase基因序列,构建了aldolase/pGEX-4T-1原核表达系统,表达并纯化了谷胱苷肽巯基转移酶-醛缩酶(GST-aldolase)融合蛋白。结论体外获得了纯化的GST-aldolase蛋白,为后续的aldolase功能研究奠定了基础。更多还原

【Abstract】 Objective To obtain and purify the aldolase protein of Toxoplasma gondii in vitro.Methods The aldolase gene was obtained from cDNA library by polymerase chain reaction amplification,and subcloned into pGEX-4T-1 to generate translational fusion with glutathione-S-transferases(GST).The recombinant protein GST-aldolase was expressed as a fusion protein with a GST tag in E.coli upon isopropyl-β-D-thigalactopyranoside induction and then purified with affinity chromatography.Results The aldolase gene was obtained,and the recombinant plasmid aldolase/pGEX-4T-1 was constructed successfully and expressed as a fusion protein.Conclusion The protein GST-aldolase is expressed in vitro,which may provide the foundation for the further studies on the function of aldolase.更多还原