CAJ | 학술논문

目的：构建针对人信号传导及转录激活因子1（STAT1）基因的shRNA表达载体，检测其对STAT1基因的沉默效果。方法设计针对STAT1的shRNA序列，克隆到pGPU6质粒载体，并进行测序、转染和鉴定。结果 DNA测序显示成功构建了4个shRNA表达载体。shRNA能有效抑制STAT1基因在人宫颈癌细胞株TZM- bl细胞中的表达，抑制率为63%。结论成功构建了针对人STAT1基因的shRNA表达载体pGPU6/GFP/Neo- shRNA STAT1。
목적：구건침대인신호전도급전록격활인자1（STAT1）기인적shRNA표체재체，검측기대STAT1기인적침묵효과。방법설계침대STAT1적shRNA서렬，극륭도pGPU6질립재체，병진행측서、전염화감정。결과 DNA측서현시성공구건료4개shRNA표체재체。shRNA능유효억제STAT1기인재인궁경암세포주TZM- bl세포중적표체，억제솔위63%。결론성공구건료침대인STAT1기인적shRNA표체재체pGPU6/GFP/Neo- shRNA STAT1。
Objective To construct pGPU6 shRNA expression vector targeting human STAT1 gene and to evaluate its si-lencing effect in TZM- bl cellline. Methods The targeting sequences were designed and synthesized, and cloned into the pG-PU6/GFP/Neo vector. After confirmation by DNA sequencing, the recombinant vectors were transfected into TZM- bl cells, and the expression levels of STAT1 were determined. Results Four shRNA clones were successful y constructed and confirmed by DNA sequencing. The transfected cells presented gene silencing effect and the protein expression of STAT1 was down- regulat-ed by 63%. Conclusion We have successful y constructed STAT1- shRNA expression vector and its effect is validated in trans-fected TZM- bl cells.