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目的:构建金黄色葡萄球菌肠毒素A(SEA)和表皮生长因子(EGF)表达载体。方法:利用PCR及RT-PCR技术分别克隆出SEA基因及EGF基因片断,以经过改良优化的桥式PCR将2个基因融合,再转入表达载体pET-44,经诱导剂诱导后分泌SEA-EGF融合蛋白。结果:所得SEA、EGF基因测序结果示与GENEBANK中公布的标准序列一致,且成功融合SEA-EGF基因并成功导入表达载体。结论:该研究成功构建了SEA-EGF表达载体,为进一步研究SEA-EGF融合蛋白抗头颈肿瘤靶向免疫治疗奠定了基础。
Objective: To construct Staphylococcus aureus enterotoxin A (SEA) and epidermal growth factor (EGF) expression vector. Methods: The SEA and EGF gene fragments were cloned by PCR and RT-PCR respectively. Two genes were fused by modified PCR and then transferred into expression vector pET-44. The SEA- EGF fusion protein. Results: The sequencing results of SEA and EGF genes were consistent with those published in GENEBANK. The SEA-EGF gene was successfully fused and successfully transfected into the expression vector. Conclusion: The study successfully constructed SEA-EGF expression vector, which laid the foundation for further study of targeted immunotherapy against SEA-EGF fusion protein in head and neck cancer.