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目的：构建含人白细胞介素－２（ｈＩＬ－２）基因的痘菌病毒真核表达载体ｐＭＪ６０１，为进一步实施胃癌的基因治疗作必要准备。方法：利用质粒抽提、琼脂糖凝胶电泳、酶切、目的基因与载体的连接、感受态细胞的制备及转化和ＤＮＡ序列分析等多种基因工程技术，将纯化回收的ｈＩＬ－２基因分别与质粒ｐＢｌｕｅｓｃｒｉｐｔⅡＳＫ＋／－及ｐＭＪ６０１进行连接、转化并签定。结果：克隆有ｈＩＬ－２基因的ｐＬＸＳＮ经ＥｃｏＲＩ－ＢａｍＨＩ酶切后，ｈＩＬ－２基因被成功地连接到质粒ｐＢｌｕｅｓｃｒｉｐｔⅡＳＫ＋／－上，再经Ｓａｉｌ－ＢａｍＨＩ酶切后，ｈＩＬ－２基因又被克隆到ｐＭＪ６０１真核表达载体上。结论：重组ｈＩＬ－２基因痘苗病毒真核表达载体ｐＭＪ６０１的构建为胃癌的免疫基因治疗打下了坚实的基础。
Objective: To construct eukaryotic expression vector pMJ601 containing human interleukin-2 (hIL-2) gene and provide necessary preparation for further gene therapy of gastric cancer. Methods: The purified hIL-2 gene was recovered by plasmid extraction, agarose gel electrophoresis, enzyme digestion, ligation of the target gene and vector, preparation and transformation of competent cells, and DNA sequence analysis. The plasmid pBluescriptIISK+/- and pMJ601 were ligated, transformed and signed. Results: pLXSN cloned with hIL-2 gene was digested with EcoRI-BamHI. The hIL-2 gene was successfully ligated into plasmid pBluescriptIISK+/-. After being digested with Sail-BamHI, the hIL-2 gene was cloned. pMJ601 eukaryotic expression vector. Conclusion: The construction of recombinant eukaryotic expression vector pMJ601 of hIL-2 gene vaccinia virus lays a solid foundation for the immunogenic therapy of gastric cancer.