HER-2模拟表位mut串联肽载体的构建及表达  

Construction and Expression of Repeats of HER-2 Mimetic Peptide

作　　者：赵丽丽[1,2,3] 孟麟[1,2,3] 姜北海[1,2,3] 

机构地区：[1]北京大学临床肿瘤学院 [2]北京肿瘤医院 [3]北京市肿瘤防治研究所生化与分子生物学实验室恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142

出　　处：《生物医学工程学杂志》2010年第5期1076-1079,1099,共5页Journal of Biomedical Engineering

基　　金：国家"863"计划项目资助(2007AA021103)

摘　　要：应用HER-2 B细胞表位模拟小肽mut,构建含有该小肽的原核串联肽表达载体,使表达蛋白产生多个小肽抗原表位。将mut小肽DNA序列定向插入pET28a(+)载体中,构建原核表达载体pET28a-mut1,在此基础上将mut基因序列进行同向串联,获得一系列串联肽表达载体pET28a-mut2、pET28a-mut4、pET28a-mut8及pET28a-mut16,将串联肽表达载体转化大肠杆菌BL21,经常规IPTG诱导后,正确表达目的蛋白。Western blot结果提示,mut小肽经串联后,可增强与Heceptin的结合能力。小肽串联表达可产生多个抗原表位,从而为进一步小肽疫苗的功能研究及HER-2疫苗的研制奠定了一定基础。To produce many epitopes of peptide mut,different tandem repeats containing HER-2 mimetic peptide mut were constructed and expressed.The oligonucleotide coding mut sequence was inserted into pET28a（＋） to construct the prokaryotic recombinant plasmid pET28a-mut1.The mut coding sequence was repeatedly inserted into the vector in tendem to obtain a series of plasmids pET28a-mut2,pET28a-mut4,pET28a-mut8 and pET28a-mut16.The plasmids were transformed into E.coli.BL21 and induced by IPTG.The recombinant proteins were expressed in E.coli.The binding analysis verified that the interaction between the tandem peptides mut and Herceptin was increased.In conclusion,the tandem peptides consisted of many antigen epitopes,and laid the foundation for the further study of HER-2 peptide vaccine for biotherapy of cancer with HER-2 overexpression.

关 键 词：HER-2 串联肽 疫苗 

分 类 号：R346[医药卫生—基础医学]