寡聚蛋白Trim34α对NF-κB报告基因的负向调节作用

目的 测Trim34α对TAB2诱导的NF-κB报告基因活化的影响.结果 经鉴定成功构建Trim34α真核表达载体,该载体表达的Trim34α蛋白能相互聚集形成寡聚体(Trim小体);Trim34α可显著抑制TAB2诱导的NF-κB荧光素酶报告基因活化.结论 Trim34α可以在胞内形成寡聚体,Trim34α能显著抑制TAB2诱导的NF-κB报告基因活化.

Abstract：

Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.