两种人源性doc1基因真核表达重组质粒的构建及表达

Construction of the eukaryotic recombinant plasmid of doc1 gene of human

目的:构建p12蛋白真核表达重组载体,为基因治疗口腔癌提供基础研究。方法:反转录-聚合酶链式反应克隆出doc1基因ORF读框,两端连入酶切位点,导入真核表达载体pT7-FLAG-4及pEGFP-N1。结果:重组子pT7-FLAG-4-doc1及pEGFP-N1-doc1经限制性酶切分析,PCR鉴定和序列测定为正确重组质粒。结论:成功构建p12蛋白真核表达重组质粒pT7-FLAG-4-doc1及pEGFP-N1-doc1,为今后瞬时和稳定转染细胞提供基础。

Objective：To construct recombinant plasmid pTT-FLAG-4-doc1 and pEGFP-N1- doc1,so as to provide basis for gene treatment of oral cancer. Method：The doc1 gene was obtained from HaCAT cell by PCR and was subcloned into eukaryotic expressed vector pTT-FLAG-4 and pEGFP-N1- doc1.The recombinant plasmids were named pTT-FLAG-4-doc1 and pEGFP-N1- doc1 ,were identified by restriction analysis,PCR and DNA sequencing analysis. Result：The recombinant plasmid pT7-FLAG-4-doc1 and pEGFP-N1-doc1were identified by restriction analysis, PCR and DNA sequencing analysis. Conclusion： The recombinant plasmid pTT-FLAG-4-doc1 and pEGFP-N1- doc1 were successfully constructed.