利用RNAi技术抑制ARPC2基因表达的研究  

Inhibition of gene expression of ARPC2 by RNAi

作　　者：龚小卫[1] 彭毅[1] 刘靖华[1] 邓鹏[1] 刘志锋[1] 秦清和[1] 姜勇[1] 

机构地区：[1]广州南方医科大学病理生理学教研室,510515

出　　处：《解放军医学杂志》2004年第10期882-884,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家杰出青年科学基金 (编号 3992 50 1 4 );国家重大基础理论研究计划(编号 2 0 0 2CB51 30 0 0 )资助课题

摘　　要：目的　构建pSUPER ARPC2表达载体 ,并在真核细胞中进行表达 ,验证ARPC2基因表达是否受到抑制。方法　设计特异性针对ARPC2基因的寡核苷酸序列 ,退火后利用常规酶切、连接方法将其重组至pSUPER .basic载体中。对阳性克隆进行酶切和测序鉴定后 ,转染HEK 2 93细胞 ,利用RT PCR和Westernblot检测ARPC2基因的表达情况。结果　构建的pSUPER ARPC2载体导入HEK 2 93细胞时 ,ARPC2基因的表达受到抑制 ,蛋白合成减少。结论　成功构建了ARPC2的RNAi表达载体 ,并证实其能对ARPC2基因的表达产生抑制作用 ,为进一步研究ARPC2的生理功能提供了重要的实验材料。Objective To construct pSUPER-ARPC2 vector and to obtain its expression in eukaryotic cell, in order to down-regulate the expression of ARPC2. Methods Oligonucleotide sequences specific for ARPC2 were designed for RNAi, and cloned into pSUPER.basic vector after annealing by restriction enzyme digestion and ligation. After identification by restriction enzyme digestion and sequencing, the plasmid of positive clones was transfected into HEK 293 cells for three times. The expression of ARPC2 was detected by RT-PCR and Western blot. Results It was found that the expression of ARPC2 was definitely knocked down by transfection with pSUPER-ARPC2. Conclusions The RNAi expression vector of ARPC2 was constructed successfully, which was showed to possess the ability to knockdown the expression of ARPC2 by different methods. So, pSUPER-ARPC2 should be a powerful tool in the study of function of ARPC2.

关 键 词：基因表达 基因 抑制 HEK293细胞 转染 寡核苷酸 蛋白合成 表达载体 RNAI技术 阳性克隆 

分 类 号：R735[医药卫生—肿瘤] Q78[医药卫生—临床医学]