摘要
将 Cu,Zn- SOD与抗癌胚抗原 ( CEA)单链抗体基因 ( Sc Fv gene)融合 ,重组到含 T7启动子的表达载体 p ET- 2 2 b( + )中 ,构建表达质粒 p ETSOD- Sc Fv,并转化大肠杆菌 BL2 1 ( DE3) ,进行高效表达 ,表达物占菌体可溶性总蛋白的 1 8%。SDS- PAGE和蛋白质印迹图谱显示表达物相对分子质量为 41 k D,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。
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.
出处
《华东理工大学学报（自然科学版）》
CAS
CSCD
北大核心
2001年第2期124-127,共4页
Journal of East China University of Science and Technology
关键词
癌胚抗原
单链抗体
Cu
Zn-SOD
融合表达
基因表达
克隆
carcinoembryonic antigen
single chain antibody
copper, zinc superoxide dismutase
fusion expression
targeting
