TRAF6基因RNA干涉表达载体的构建及鉴定

Construction and identification of siRNA against tumor necrosis factor receptor associated factor 6 (TRAF6)

目的构建针对小鼠肿瘤坏死因子受体相关因子6(TRAF6)的mRNA的siRNA真核表达载体,并鉴定其正确性。方法利用基因重组技术,化学合成用于产生针对TRAF6的发卡状RNA的寡核苷酸,各64个碱基,退火后插入线性化的pSUPER载体的H1启动子下游。重组载体分别用限制性酶切和测序鉴定其正确性。结果限制性酶切和序列测定表明成功构建了可以表达针对TRAF6的发卡状RNA表达载体。结论成功构建了针对TRAF6的发卡状RNA表达载体,为研究TRAF6的功能和进一步的基因治疗提供了实验基础。

Objective To construct and identify vectors expressing siRNA against tumor necrosis factor receptor associated factor 6 （TRAF6）. Methods Using gene recombination techniques, 64-bp oligonucleotidcs for hairpin RNA targeting TRAF6 were synthesized, annealed, and inserted into the downstream of H1 promoter of linearized pSUPER to form recombinant vectors, which were identified by restriction digestion and DNA sequencing. Results Restriction digestion and DNA sequencing showed suecessihl construction of TRAF6-targeting siRNA. Conclusion Vectors ex- pressing hairpin RNA against TRAF6 were successfully produced, which would facilitate further studies of TRAF6 function and its use in geuotherapy.