重组质粒pcDNA3/sIL-1R(I)在哺乳动物细胞中的表达

Expression of Constructed Eukaryotic Vectors Carrying Encoding Gene of Soluble Human Interleukin-1 Receptor in Mammalian Cells

目的 检测重组质粒pcDNA3 sIL_1R(I)在哺乳动物细胞系COS_1和CHO细胞中的表达。方法 采用脂质体介导基因转染技术 ,将已构建的pcDNA3 sIL_1R(I)质粒DNA导入体外培养的哺乳动物细胞系COS_1和CHO细胞中。加入G4 18对转染的细胞加压筛选 ,获得稳定转染的细胞。酶联免疫吸附实验 (ELISA)对重组质粒在COS_1和CHO细胞表达产物的含量进行检测。结果 G4 18筛选获得稳定转染的COS_1和CHO细胞 ,对照组加压后第10～ 2 0天细胞全部死亡。转染组第 15～ 2 3天汇片达 80 %左右。转染组细胞培养液和冻融液中sIL_1R表达量均显著高于对照组 (P <0 0 5 )。结论 以sIL_1R胞外成熟肽编码区基因作为目的基因构建的重组质粒pcDNA3 sIL_1R(I)在哺乳动物细胞系中具有表达功能。

Objective This study is to make sure if the constructed pcDNA3 carrying encoding gene of sIL-1R can be expressed in mammalian cells in vitro.Methods COS-1 cells and CHO cells were respectively transfected with recombinant plasmid pcDNA3/sIL-1R by liposome. The protein expression products were detected by ELISA.Results The results indicated that the protein expression products could be detected in the cell plasma and the cell culture supernatant. The expression level in experimental groups was much higher than that in control groups(P<0.05).Conclusion The constructed pcDNA3/sIL-1R can express interest protein in mammalian cells and this establishes the basis for future investigation on gene therapy of periodonititis and other inflammatory diseases.