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)

作　　者：李霞[1] 肖明振[2] 余擎[2] 赵守亮[2] 李卫星[1] 罗晓晋[1] 王捍国[2] 

机构地区：[1]山西医科大学口腔医学系,太原第四军医大学博士生030001 [2]第四军医大学秦都口腔医院牙体牙髓病科

出　　处：《中国药物与临床》2007年第7期502-504,共3页Chinese Remedies & Clinics

基　　金：国家自然科学基金资助项目(30271418);山西省青年科技开发基金资助项目(2006021041)

摘　　要：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.

关 键 词：受体 肿瘤坏死因子 基因表达 遗传载体 

分 类 号：R346[医药卫生—基础医学]