CAJ | 학술논문

在大肠杆菌中表达小鼠金属硫蛋白-I，并研究其分子结构和生物学功能。方法：将小鼠金属硫蛋白-I基因克隆入融合表达载体pGEX-4T-1中，转化大肠杆菌BL21，得到含重组表达质粒pGMA的工程菌。结果：经IPTG诱导，表达出分子量约为33KD的融合蛋白。然后经Glutathione-Sepharose 4B亲和纯化后，用凝血酶切除GST部分。经Sephacryl S-100过滤纯化，得到结合有Cd2+的小鼠金属硫蛋白-I。产率约为3mg/L培养液。结论：成功制备了表达MT的工程菌。
재대장간균중표체소서금속류단백-I，병연구기분자결구화생물학공능。방법：장소서금속류단백-I기인극륭입융합표체재체pGEX-4T-1중，전화대장간균BL21，득도함중조표체질립pGMA적공정균。결과：경IPTG유도，표체출분자량약위33KD적융합단백。연후경Glutathione-Sepharose 4B친화순화후，용응혈매절제GST부분。경Sephacryl S-100과려순화，득도결합유Cd2+적소서금속류단백-I。산솔약위3mg/L배양액。결론：성공제비료표체MT적공정균。
To express mouse metallothionein-I in Escherichia coli and study its molecular structure and function. Methods:Gene encoding the mouse MT-I was cloned into fusion vector pGEX-4T-1 and transformed into Escherichia coli BL21. Results:With IPGT inducing,a fusion protein whose molecular weight was about 33KD was expressed. The fusion protein was purified through Glutathione-Sepharose 4B affinitive chromatography and GST part was cut off with thrombin. Finally, the mixture was purified through Sephacryl S100 gel filtration and the purified MT-I protein binding Cd2+ was obtained. The productivity is 3mg/L cultrure. Conclusion:The recombinant MT expression vector was successfully constructe.