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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。
The HER-2 B cell epitope was used to mimic the small peptide mut, and a prokaryotic tandem peptide expression vector containing the small peptide was constructed to make the expressed protein produce a plurality of small peptide epitopes. The small peptide sequence of mut was inserted into pET28a (+) vector and the prokaryotic expression vector pET28a-mut1 was constructed. Based on the mutagenesis, the mut was cloned into the vector pET28a-mut2 and pET28a-mut2 , PET28a-mut8 and pET28a-mut16. The tandem peptide expression vector was transformed into E.coli BL21 and expressed by IPTG. Western blot results suggest that, mut peptide in series, can enhance the binding capacity with Heceptin. Small peptide tandem expression can produce multiple epitopes, which will lay the foundation for the further study of the function of small peptide vaccine and the development of HER-2 vaccine.