靶向性超抗原EGF-SEA融合基因的构建及表达

Construction and expression of the targeting super-antigen EGF-SEA fusion gene

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 expression vector for the SEA-EGF fusion gene. Method： Clone the SEA gene and the EGF gene segment with PCR and RT-PCR independently, and connect this two genes by the bridge PCR. Insert the fusion gene EGF-SEA into the expression vector PET-44. Induced the secretion of the fusion protein SEA-EGF by the antileptic. Result：The gene fragment encoding EGF and SEA mature peptide was successfully cloned. The fusion gene EGF-SEA was successfully constructed and was inserted into expression vector. Conclusion：The new recombinant expression vector for fusion gene EGF-SEA is specific for head and neck cancer,laid the foundation for the further study of fusion protein SEA-EGF targeting immune therapy in head and neck tumors.