Cloning and expression of murine TLR2 gene and study on its biological activity

The Toll-like receptor 2(TLR2) gene was amplified from mouse macrophages by RT-PCR.After being cloned into the pMD18-T vector,TLR2 gene was subcloned into the pcDNA3.1 vector to construct the eukaryotic expression plasmid pcDNA3.1-mTLR2.The recombinant plasmid was transiently transfected into the HEK293T cells, its expression and subcellular localization were analyzed by using RT-PCR and immunofluorescent assay,respectively. The results indicated that mTLR2 was successfully transfected,expressed,and localized at the cell membrane.TLR2 positive activator pam3csk4 was used to stimulate the transfected HEK293T cells,dual luciferase report gene system was applied to measure the transcription activity of NF-κB.The luciferase activity of the group stimulated by pam3csk4 was significantly higher than that of the normal saline control group,implying the recombinant mTLR2 had wild type function. In conclusion, the mTLR2 gene was successfully cloned and expressed and the protein had biological activity.This study provided the base for studying on TLR2 signal pathway and its role in resistance to parasite infection.