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

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.