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
作者：徐艳,李鹏,张蕴惠,章锦才,王大章
Author：XU Yan*,LIPeng,ZHANG Yunhui,et al.
收稿日期：2003-12-25 年卷（期）页码：2003,21(06):467-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：人可溶性白细胞介素-1受体,哺乳动物细胞,体外转染,基因治疗,
Key words：human soluble interleukin-1 receptor,mammalian cells,in vivotransfer,gene therapy,
基金项目：
本课题为国家自然科学基金(编号3917081)及江苏省自然科学基金(编号2003422)资助项目
中文摘要
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G418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 G418筛选获得稳定转染的COS-1和CHO细胞,对照组加压后第 10~20天细胞全部死亡。转染组第15~23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
英文摘要
Objective This study is to make sure if the constructed pcDNA3 carrying encoding gene of sIL-1R can be expressed in mammalian cellsin 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
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