pEGFP-N1-PEDF重组质粒的构建和鉴定

Construction and identification of eukaryotic expression vector pEGFP-N1-PEDF

目的构建pEGFP-N1-PEDF真核表达质粒,并进行鉴定。方法利用RT-PCR方法从组织中提取PEDF的基因片段,与pMD18-T载体连接,构建pMD18-T-PEDF重组质粒,酶切后与pEGFP-N1真核表达载体连接,构建pEGFP-N1-PEDF真核表达质粒,进行测序、酶切鉴定,表明质粒构建成功。结果本实验成功构建了pEGFP-N1-PEDF真核表达质粒。结论为进一步研究利用PEDF基因治疗皮肤肿瘤提供实验基础。

Objective To construct an eukaryotic expression vector carrying PEDF gene. Methods PEDF gene was cloned from tissue by using RT-PCR method and was inserted into pMD18-T to construct pMD18-T-PEDF plasmid ,and to construct pEGFP-N1-EDF plasmid by using restriction enzyme.To identify the plasmid by sequencing and restriction enzyme. Results The eukaryotic expression vector pEGFP-N1-PEDF was constructed and identified. Conclusion It provides the base of apply the PEDF in treatment of skin tumor.