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目的:在大肠杆菌中表达、并纯化获得高纯度的融合蛋白疫苗HSP65-HER2。方法:经过酶切和序列分析后,将pET-28a-HSP65-HER2重组质粒转化Ecoli(BL21)中,经IPTG诱导产生6-his tag-HSP65-HER2融合蛋白,并通过Ni Sepharose 4B和Superdex G-25纯化。以纯化的重组融合蛋白免疫小鼠,取脾细胞经体外诱导后,采用CTL活性检测其活性。结果:成功的表达、纯化获得了高纯度的HSP65-HER2融合蛋白,CTL活性检测结果表明具有生物学活性。结论:获得了高纯度的HSP65-HER2融合蛋白,并证明其具有免疫学活性,为进一步研究其生物学活性奠定了基础。
OBJECTIVE: To express in Escherichia coli and obtain high-purity fusion protein vaccine HSP65-HER2. Methods: The recombinant plasmid pET-28a-HSP65-HER2 was transformed into Ecoli (BL21) after digestion and sequence analysis. The 6-his tag-HSP65-HER2 fusion protein was induced by IPTG and purified by Ni Sepharose 4B and Superdex G -25 purification. The purified recombinant fusion protein was used to immunize mice. After spleen cells were induced in vitro, their activity was detected by CTL activity. RESULTS: High purity HSP65-HER2 fusion protein was successfully expressed and purified. The results of CTL activity showed that it has biological activity. CONCLUSION: High-purity HSP65-HER2 fusion protein was obtained and proved to be immunologically active, which laid the foundation for further study of its biological activity.