SV40LT基因过表达慢病毒载体的构建与包装

Construction of Lentiviral Vector Overexpressing SV40LT

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p Lenti-GFP质粒中,构建重组穿梭质粒p Lenti-GFP-SV40LT,测序鉴定后分别将鉴定的阳性p Lenti-GFP-SV40LT和包装质粒p MD2.0G和ps PAX2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。

Objective To construct a lentiviral vector overexpressing of SV40 LT,which serve a base to establish the immortalization of embryonic fibroblasts. Method SV40 LT gene was obtained from 293 T cells and then inserted into p Lenti-GFP vector to construct recombinant plasmid p Lenti-GFP-SV40 LT,which identified by nucleotide sequencing. p Lenti-GFP-SV40 LT, p MD2. 0G, ps PAX2 were co-transfected into 293 T cells for lentivirus packaging. Result The Construction of Lentiviral Vector Over-expressing SV40 LT was constructed successfully.Conclusion The recombinant lentivirus carrying SV40 LT gene was constructed successfully, which lay a foundation to study immortalization of embryonic fibroblasts.