携带三种报告基因慢病毒载体的构建及其实验研究  

Construction of a Lentivirus Vector Expressing Three Reporter Genes and Its Experimental Study

作　　者：王丽娴[1] 王东阳[1] 李星[1] 夏海滨[1] 

机构地区：[1]陕西师范大学生命科学学院基因治疗研究室,西安710062

出　　处：《中国生物工程杂志》2011年第12期86-92,共7页China Biotechnology

基　　金：国家自然科学基金资助项目(3087299331070137)

摘　　要：慢病毒是逆转录病毒科的一个属。它具有可以感染分裂期及非分裂期细胞、容纳外源性目的基因片段大、目的基因表达时间较长、免疫反应小等诸多优点,因此成为运载目的基因的理想载体而得到广泛的应用。构建了带有3种报告基因即红色荧光蛋白(mCherry)、荧光素酶基因(luciferase)及绿色荧光蛋白(eGFP)的慢病毒载体,其中mCherry及eGFP由于其成像的直观性特点便于用于体外细胞株的筛选,而luciferase由于其敏感性高,则可以用于灵敏检测动物体内肿瘤模型的生长及转移情况。选择了一种恶性程度较高的脑胶质瘤母细胞U87MG(malignantglioblastoma)作为研究对象,体外成功构建了表达3种报告基因的细胞株并建立了裸鼠体内肿瘤模型,为该肿瘤疾病动物体内的实验治疗效果评估及肿瘤转移的检测奠定了基础;同时也为携带报告基因的其它肿瘤细胞系及其肿瘤模型的建立提供了方便的载体工具。Lentiviral is a genus of retroviridae, which has minor immune response and can deliver and express large exogenous gene fragment for longer time. Therefore, it is widely used as an ideal vector for delivering the target gene. One type of lentivirus vector containing three types of reporter gene, e.g. mCherry, luciferase and eGFP was constructed. Mcherry and eGFP can be used for cell line screening in vitro because of its intuitive imaging, while luciferase can be used for detecting the growth and metastasis of the tumor cells on animal models in vivo sensitively. The constructed vector was transfected to the malignant glioblastoma U87MG successfully, and the corresponding animal model was developed on nude mice by planting the transfected cells. It provided a model for the evaluation of therapeutic effect on glioblastoma in vivo, and demonstrated a reference way for other tumor cell lines and models which carried reporter genes

关 键 词：慢病毒载体 报告基因 脑胶质瘤 动物模型 

分 类 号：Q789[生物学—分子生物学]