摘要
目的:构建CDK2的干扰RNA真核表达载体,并且稳定转染人脑胶质细胞瘤SHG44细胞来抑制CDK2的表达,为人脑胶质细胞瘤的研究提供有价值的资料。方法:1.根据siRNA设计原则和GeneBank数据库中CDK2的cDNA序列,构建CDK2干扰RNA真核表达载体PGenesil-1-CDK2,并测序鉴定。2.利用脂质体法转染CDK2的干扰RNA真核表达载体,G418筛选阳性转染细胞克隆,制备稳定转染干扰RNA真核表达载体的SHG44细胞系,用倒置荧光显微镜观察荧光蛋白表达量。结果:1.成功构建了CDK2干扰RNA真核表达载体。经鉴定证实,构建的siRNAs序列与基因库中序列完全相同,并且未发现有突变、缺失、插入等异常存在。2.获得稳定转染CDK2干扰RNA真核表达载体的SHG44细胞系,命名为PGenesil-1-CDK2-SHG44。结论:成功的建立稳定转染CDK2干扰RNA的SHG44细胞系。
Objective：To explore the function of CDK2,a stable expression of CDK2 SiRNA astrocytoma cell line was established.Method： The Eukaryotic expression vector PGenesil-1-CDK2,CKD2 specific RNA interference,was constructed based on the sequence from Genbank.The plasmid was sequenced and transfected to SHG44 cell line using oligofectamine.The stable transfectants were selected by G418.Results： The sequence of the construct was confirmed right.All stable transfectants had significant lower expression of CDK2 compared with the control clones.Conclusion： The PGenesil-1-CDK2-SHG44 stable transfectants were successfully established with the constructed PGenesil-1-CDK2vector.
出处
《内蒙古医学院学报》
2011年第6期469-472,共4页
Acta Academiae Medicinae Neimongol
基金
内蒙古医学院附属医院博士启动金资助课题
