CAJ | 학술논문

将Cu,Zn-SOD与抗癌胚抗原(CEA)单链抗体基因(ScFv gene)融合，重组到含T7启动子的表达载体pET-22b(+)中，构建表达质粒pETSOD-ScFv，并转化大肠杆菌BL21(DE3)，进行高效表达，表达物占菌体可溶性总蛋白的18%。SDS-PAGE和蛋白质印迹图谱显示表达物相对分子质量为41kD，与融合基因编码蛋白质的理论值相符。该蛋白质在大肠杆菌中为分泌型表达，有利于纯化。RIA测定表明表达产物能特异性地与抗原CEA结合，邻苯三酚法测定也表明表达产物具有SOD酶的活性，该融合蛋白为分泌CEA肿瘤的靶向性治疗及放疗、化疗后的恢复提供新的途径。
장Cu,Zn-SOD여항암배항원(CEA)단련항체기인(ScFv gene)융합，중조도함T7계동자적표체재체pET-22b(+)중，구건표체질립pETSOD-ScFv，병전화대장간균BL21(DE3)，진행고효표체，표체물점균체가용성총단백적18%。SDS-PAGE화단백질인적도보현시표체물상대분자질량위41kD，여융합기인편마단백질적이론치상부。해단백질재대장간균중위분비형표체，유리우순화。RIA측정표명표체산물능특이성지여항원CEA결합，린분삼분법측정야표명표체산물구유SOD매적활성，해융합단백위분비CEA종류적파향성치료급방료、화료후적회복제공신적도경。
The gene encoding human Cu,Zn-SOD was fused to anti-carcinoembryonic antigen single-chain antibody gene to construct the fusion gene, then was ligated into prokaryotic expression vector pET-22b(+). The fusion gene was expressed in E.coli at high level, and was characterized by SDS-PAGE and Western-blot analysis. The expression product which accounting for 18% of the total bacteria protein had the CEA-binding ability in RIA, and also had the SOD activity by pyrogallol autoxidition assay. So, the Cu,Zn-SOD moiety retains substantial enzymatic activity, where the ScFv moiety can delivers the fusion protein to tumor. Cu,Zn-SOD is a potential tumor-relevant gene, maybe the fusion protein can provide a new pathway to therapy tumors and to meet the damage after radiation or chemical therapy.