MAGE-1与IL-18共表达DNA疫苗的构建与体外表达

目的构建MAGE-1与IL-18共表达质粒pcIL-18-MAGE并进行体外验证表达。方法设计合成MAGE-1与IL-18引物,PCR扩增其基因片段,分别构建以pcDNA3为载体的小鼠IL-18重组质粒和人MAGE-1重组质粒,pcCMV-IL-18与MAGE-1的PCR产物酶切连接构建共表达质粒pcIL-18-MAGE。脂质体法将构建的重组质粒转染293细胞,RT-PCR和Western印迹法验证MAGE-1和IL-18在转化细胞中的表达。结果成功获得共表达质粒pcIL-18-MAGE,RT-PCR可见目的条带,Western印迹显示转染MAGE-1与IL-18的细胞在蛋白质水平上均有表达。结论成功地构建出既表达mIL-18又表达MAGE-1的共表达质粒pcIL-18-MAGE。

Construction of a DNA vaccine co-expressing both MAGE-1 and interleukin-18 and its expression in vitro

Objective To construct a co-expressing plasmid pclL-18-MAGE of MAGE-1 and IL-18, and verify its expression in vitro. Methods MAGE-1 and IL-18 were amplified by PCR with primers, which were respectively inserted into carrying agent pcDNA3 to construct recombinant plasmids of mice IL-18 and human MAGE-1 and to connect products of pcCMV-IL-18 and MAGE-1 to obtain co-expression plasmid pcIL-18-MAGE. The recombinant plasmids were transfected into 293 cells by LipofectamineTM , and expression of MAGE-1 and IL-18 were detected by RT-PCR and Western blot. Results Co-expressing plasmid pcIL-18-MAGE was gained successfully, RT-PCR analysis of cells displayed positive bands,Western blot displayed that MAGE-1 and IL-18 expressed at protein level. Conclusions Co-expressing plasmid pcIL-18-MAGE which can express both MAGE-1 and IL-18 is constructed successfully.