SOCS2真核表达载体的构建及在3T3-L1前体脂肪细胞中的表达  

Construction and Expression in 3T3-L1 of Eukaryotic Pasmid Carrying Mouse SOCS2 Gene

作　　者：孙婵[1] 孙超[1] 解芸菲[1] 安磊[1] 王勇[1] 吕彬[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《西北农业学报》2011年第6期18-22,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金项目(30871785);教育部新世纪优秀人才计划项目(NCET20620865)

摘　　要：细胞因子信号抑制蛋白(SOCS)是细胞因子信号通路的重要负调控因子。构建SOCS2真核表达载体并转染3T3-L1前体脂肪细胞,为研究SOCS2对不同细胞因子调控脂代谢奠定基础。取小鼠皮下脂肪提总RNA进行RT-PCR,克隆获得pMDl8-SOCS2质粒载体,双酶切目的基因,并克隆至质粒载体pcDNA3.1,成功构建了重组质粒载体pcDNA3.1-SOCS2;重组质粒瞬时转染3T3-L1前体脂肪细胞系,以未转染和空载体转染作为对照,RT-PCR和Western blotting检测重组质粒转染组SOCS2核酸和蛋白表达,表达水平均显著高于对照组,为进一步研究SOCS2基因对不同细胞因子调控脂肪代谢提供基础。Suppressors of cytokine signaling（SOCS） proteins are a key group of negative feedback regulators of cytokine signaling pathway.To construct and express in 3T3-L1 of eukaryotic plasmid carrying mouse suppressors of cytokine signaling-2（SOCS-2） gene.Total RNA was extracted from mouse subcutaneous fat,plasmids of pMDl8-SOCS2 and pcDNA3.1-SOCS2 was successfully constructed after RT-PCR and TA cloned.Plasmid pcDNA3.1-SOCS2 was confirmed by restriction digestion and gene sequencing.The sequence of SOCS2 gene was consistent with that of NCBI gene bank.The recombinant plasmid pcDNA3.1-SOCS2 was transfected into 3T3-L1 cell line.The mRNA and protein of SOCS2 gene were identified by real-time PCR and western blot analysis.The non-transfected 3T3-L1 cells and 3T3-L1 cells transfected with empty vector were served as controls.The mRNA and protein of SOCS2 expression in recombinant pcDNA3.1-SOCS2 group was significantly higher than that in un-transfected group and empty vector group.Plasmid pcDNA3.1-SOCS2 was successfully constructed and expressed in 3T3-L1 cell line.

关 键 词：细胞因子信号抑制蛋白2(SOCS2) 真核表达载体PCDNA3.1 细胞因子 

分 类 号：Q26[生物学—细胞生物学]