LeftyA基因重组慢病毒表达载体的构建及鉴定  

Construction and Indentification of LeftyA Recombinant Lentiviral Expression Vector

作　　者：邱宇安[1] 陈文学[1] 靳文剑[1] 陈火国[1] 钟宁[1] 余洁丽 

机构地区：[1]江西省肿瘤医院,330029

出　　处：《实用癌症杂志》2015年第12期1755-1757,共3页The Practical Journal of Cancer

基　　金：江西省青年科学基金资助项目(编号:20132BAB215018)

摘　　要：目的想构建LeftyA基因的慢病毒表达载体,鉴定其在293T细胞中的表达。方法首先扩增LeftyA基因序列,将其与慢病毒骨架载体连接后,鉴定慢病毒表达载体构建成功。将重组慢病毒表达载体质粒及包装质粒共转染293T细胞,鉴定慢病毒转染并包装成功。结果 LeftyA基因成功转导至以GV287为骨架质粒的慢病毒系统中,包装后测定病毒滴度达2.0×108TU/ml,免疫印记法检测到LeftyA蛋白在293T细胞中的表达。结论成功构建了携带人LeftyA基因的慢病毒表达系统。Objective To construct LeftyA gene lentiviral vector system and determine its expression in 293Tcells. Methods Full-length sequence of LeftyA was amplified by PCR,and was connected with lentiviral vector.Gene sequencing was used to identify that recombinant lentiviral vector was constructed successfully.The plasmids were transfected into 293Tcells.In-dentification by fluorescent microscopy confirmed lentiviral transfection and packaging successfully.Results LeftyA were sub-cloned into GV287 as skeleton plasmid for a lentivirus packing system.The virus titer was 2.0 ×10^8 TU/ml in the supernatant liq-uid.The proteins of LeftyA were detected by western blot.Conclusion The LeftyA gene recombinant lentiviral expression system is constructed successfully.

关 键 词：LeftyA 慢病毒载体 

分 类 号：R73-36[医药卫生—肿瘤]