SV40LT 基因过表达慢病毒载体的构建与包装

摘要: 目的构建SV40LT 基因过表达慢病毒载体，并对其进行慢病毒包装，为建立永生化的uncv 小鼠胚胎成纤维细胞奠定基础。方法从293T 细胞中获得SV40LT 基因，将其克隆到pLenti-GFP 质粒中，构建重组穿梭质粒pLenti-GFP-SV40LT，测序鉴定后分别将鉴定的阳性pLenti-GFP-SV40LT 和包装质粒pMD2. 0G 和psPAX2 共转染293T 细胞，包装产生慢病毒。结果SV40LT 基因过表达慢病毒载体的构建与包装成功。结论SV40LT 基因过表达慢病毒载体构建与包装的成功为uncv 小鼠胚胎成纤维细胞的永生化提供了工具。

Construction of Lentiviral Vector Overexpressing SV40LT

Abstract: Objective To construct a lentiviral vector overexpressing of SV40LT，which serve a base to establish the immortalization of embryonic fibroblasts． Method SV40LT gene was obtained from 293T cells and then inserted into pLenti-GFP vector to construct recombinant plasmid pLenti-GFP-SV40LT，which identified by nucleotide sequencing． pLenti-GFP-SV40LT， pMD2. 0G， psPAX2 were co-transfected into 293T cells for lentivirus packaging． Ｒesult The Construction of Lentiviral Vector Over-expressing SV40LT was constructed successfully．Conclusion The recombinant lentivirus carrying SV40LT gene was constructed successfully，which lay a foundation to study immortalization of embryonic fibroblasts．