新CD44剪接变异体siRNA表达载体构建及其稳定转染5-8F细胞系的建立

发布时间：2018-04-08 17:57

本文选题：新CD44剪接变异体　切入点：siRNA载体构建　出处：《桂林医学院》2011年硕士论文

【摘要】：目的:构建CD44新剪接变异体siRNA表达载体质粒,获得质粒稳定转染的人鼻咽癌5-8F细胞系,为研究CD44新剪接变异体在鼻咽癌中的功能提供实验基础。方法:将合成的寡核苷酸链退火形成双链,连接入pGenesil-1.3-kan/Neo表达载体,并测序鉴定。脂质体法把重组质粒转染至5-8F细胞,通过G418筛选,滤纸法获取单克隆细胞,建立稳定转染的5-8F细胞系。结果:构建了新CD44剪接变异体siRNA重组真核表达载体,获得了G418筛选后的单克隆转染5-8细胞株。结论:成功构建了新CD44剪接变异体siRNA真核表达载体,成功转染到5-8F细胞内,并获得了单克隆细胞株。
[Abstract]:Aim: to construct the expression vector plasmid of CD44 splicing variant siRNA and obtain the stable transfected human nasopharyngeal carcinoma (NPC) 5-8F cell line, and to provide experimental basis for studying the function of CD44 splicing variant in nasopharyngeal carcinoma (NPC).Methods: the synthesized oligonucleotide chains were annealed to form double strands and ligated into pGenesil-1.3-kan/Neo expression vector and identified by sequencing.The recombinant plasmid was transfected into 5-8F cells by liposome method. Monoclonal cells were obtained by G418 screening and filter paper method. The stable transfected 5-8F cell lines were established.Results: the recombinant eukaryotic expression vector of new CD44 splicing variant siRNA was constructed, and the 5-8 cell line was transfected by G418.Conclusion: the eukaryotic expression vector of new CD44 splicing variant siRNA was successfully constructed and transfected into 5-8F cells successfully, and a monoclonal cell line was obtained.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.63

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