Zhao Lili, Meng Lin, Jiang Beihai
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Biochemistry and Molecular Biology, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing 100142, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2010 Oct;27(5):1076-9, 1099.
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-mutl. The mut coding sequence was repeatedly inserted into the vector in tandem 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.
为了产生肽突变体（peptide mut）的多个表位，构建并表达了包含HER-2模拟肽突变体的不同串联重复序列。将编码突变体序列的寡核苷酸插入pET28a(+)中，构建原核重组质粒pET28a-mut1。将突变体编码序列串联重复插入载体中，获得一系列质粒pET28a-mut2、pET28a-mut4、pET28a-mut8和pET28a-mut16。将这些质粒转化到大肠杆菌BL21中，并用异丙基-β-D-硫代半乳糖苷（IPTG）诱导。重组蛋白在大肠杆菌中表达。结合分析证实，串联肽突变体与赫赛汀之间的相互作用增强。总之，串联肽由许多抗原表位组成，为进一步研究用于HER-2过表达癌症生物治疗的HER-2肽疫苗奠定了基础。
