中国兽医科学2013，Vol.43Issue(3)：315-319,5.
小鼠TLR2的克隆表达及生物学活性分析
Cloning and expression of murine TLR2 gene and study on its biological activity
摘要
Abstract
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. l-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.关键词
Toll样受体2/真核表达/双荧光素酶/核转录因子-κBKey words
Toll-like receptor 2/eukaryotic expression/dual luciferase/NF-κB分类
生物科学引用本文复制引用
俞昭旸,李巍,唐颖,李兴超,刘畅,禹洋,徐佳,宋铭忻..小鼠TLR2的克隆表达及生物学活性分析[J].中国兽医科学,2013,43(3):315-319,5.基金项目
"十一五"国家科技支撑计划重大项目(2010BAD04B01) （2010BAD04B01）
国家自然科学基金资助项目(31172312) （31172312）
