CAJ | 학술논문

目的:构建携带KIR2DS2基因短发夹状干扰RNA(short hairin RNA,shRNA)的RNAi慢病毒载体.方法:针对已经筛选确定的KIR2DS2基因RNAi有效靶序列,合成靶序列的Oligo DNA,退火形成双链DNA,与经HpaⅠ和XhoⅠ酶切后的pSicoR-GFP载体[含U6启动子和绿色荧光蛋白(GFP)]连接产生LV-shKIR2DS2慢病毒载体,PCR筛选阳性克隆,测序鉴定.用LV-shKIR2DS2及Packaging system质粒共转染293T细胞,包装产生慢病毒颗粒,以293T细胞GFP蛋白的表达水平测定病毒滴度.结果:PCR和测序证实,成功构建KIR2DS2shRNA的慢病毒载体LV-shKIR2DS2.293T细胞测定病毒滴度为6×108TU/ml.结论:成功构建携带KIR2DS2基因短发夹状干扰RNA的RNAi慢病毒载体.
목적:구건휴대KIR2DS2기인단발협상간우RNA(short hairin RNA,shRNA)적RNAi만병독재체.방법:침대이경사선학정적KIR2DS2기인RNAi유효파서렬,합성파서렬적Oligo DNA,퇴화형성쌍련DNA,여경HpaⅠ화XhoⅠ매절후적pSicoR-GFP재체[함U6계동자화록색형광단백(GFP)]련접산생LV-shKIR2DS2만병독재체,PCR사선양성극륭,측서감정.용LV-shKIR2DS2급Packaging system질립공전염293T세포,포장산생만병독과립,이293T세포GFP단백적표체수평측정병독적도.결과:PCR화측서증실,성공구건KIR2DS2shRNA적만병독재체LV-shKIR2DS2.293T세포측정병독적도위6×108TU/ml.결론:성공구건휴대KIR2DS2기인단발협상간우RNA적RNAi만병독재체.