新型MUC1黏蛋白模拟表位原核表达、纯化和鉴定  

Prokaryotic Expression,Purification and Identification of New MUC1 Mimic Epitope

作　　者：崔志刚[1] 张立新[2] 岳明[1] 路浩军[2] 刘志清[2] 李春海[2] 

机构地区：[1]解放军第306医院泌尿外科,北京100101 [2]军事医学科学院附属医院肿瘤生物学实验室,北京100850

出　　处：《实用临床医药杂志》2007年第2期4-7,共4页Journal of Clinical Medicine in Practice

基　　金：国家"863"课题资助项目(2002AA214111)

摘　　要：目的对新筛选的MUC1模拟表位进行克隆表达、纯化和鉴定。方法从噬菌体随机12肽库中筛选出新的MUC1的模拟表位,目的基因片段克隆入表达质粒PET-31b(+),转化大肠杆菌BL21(DE3)plysS,IPTG诱导表达,亲和层析纯化,Western blot鉴定。结果成功构建了重组表达质粒,诱导表达出新的MUC1模拟表位重组蛋白,纯化的目的蛋白在SDS-PAGE上呈现特异的单一条带,Western印迹也检测到目的蛋白特异条带。结论成功获得了新的MUC1模拟表位表达产物,为其在肿瘤疫苗方面的应用研究创造条件。Objective To clone, express, purify and indentify a new screened MUC1 antigen mimic epitope. Methods A new MUC1 mimic epitope was screened from phage display peptide library. The gene was constructed into PET-31b（ ＋ ） expression vector and expressed in Escharichia coli BL21（DE3）plysS after transformation and induction by IPTG. The complete protein of the host bscteria was extracted for SDS-PAGE. Afterwards the recombinant protein was purified by affinity chromatography on a Ni2 ＋ - sephrose column and detected by Western blotting. Results The recombinant expression vector PET- 31b（ ＋ ） - MUC1 was constructed successfully. The fusion pro- tein was induced with IPTG and a specific protein band was shown on SDS-PAGE profile and detected by Western blotting. Conclusion The new MUC1 mimic epitope fusion protein was successfully expressed and it might be used in the development of tumor vaccines targeting MUC1.

关 键 词：MUC1 模拟表位 克隆 表达 

分 类 号：R730.5[医药卫生—肿瘤]