鼠白细胞介素-12双亚基共表达质粒的构建及在体内外的表达  

Construction of bi-cistronic co-expression plasmid of mIL-12 and the expression in vitro or in vivo

作　　者：陈玮[1] 邵传森[1] 沈建根[1] 鲍建芳[1] 潘建平[1] 韩伟[1] 寿林 项哨[1] 郑树[1] 

机构地区：[1]浙江大学医学院,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2002年第1期15-18,共4页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省卫生厅资助项目

摘　　要：目的 :构建双亚基共表达鼠白细胞介素 - 12 (m IL- 12 )真核表达质粒 ,并观察其在体内外的表达。方法 :将m IL - 12 p35和 p4 0全长编码 c DNA构建在 pc DN A 3.1载体上 ,然后把 p35表达单元 (CMV- p35 - BGH PA)插入pc DNA 3.1/ p4 0载体 ,使两个目的基因均受各自的启动子 CMV控制 ,构建成 m IL - 12双亚基共表达质粒 p Cm IL -12 ,并进行体内外表达。结果 :p Cm IL - 12在体外转染 COS- 7细胞后 ,经 EL ISA证实有 m IL- 12表达 ,其表达上清能在体外明显增强小鼠 NK细胞活性。小鼠皮内注射 p Cm IL - 12亦能增强小鼠 NK细胞活性。结论 :所构建的质粒在体内外均能表达有生物学活性的 m IL-Objective: To construct a bi cistronic co expression plasmid for mouse interleukin 12 and to observe its expression in vitro or in vivo .Methods: The full length cDNA encoding p35 and p40 was cloned into eukaryotic cells expression vector pcDNA 3.1 respectively. Subsequently, the p35 expression unit was inserted into pcDNA 3.1 /p40 to produce the bi cistronic co expression plasmid in which the p35 and p40 genes were controlled by their own CMV. The plasmid was expressed in vitro and in vivo .Results: The mIL 12 in the supernatant was detected by ELISA after the pCmIL 12 was transfected into COS 7 cells. The activity of NK cells could be augmented by the supernatant in vitro and also by by intradermal delivery of pCmIL 12 in vivo.Conclusion: The plasmid constructed by us can express biologically active mIL 12 in vitro and in vivo .

关 键 词：白细胞介素-12 生物合成 质粒 基因表达 MIL-12 天然杀伤细胞 遗传载体 分子克隆 双亚基共表达 小鼠 

分 类 号：R392[医药卫生—免疫学]