DLL1-ICD基因真核表达载体的构建及转染鉴定

通过克隆DLL1-ICD(Delta-like1细胞内区域)的cDNA及测序鉴定,构建pEGFP-C1-DLL-ICD真核表达载体并进行细胞转染,为研究Notch信号通路与Neuritin的关系奠定基础。首先根据小鼠全长DLL1 cDNA序列,设计DLL-ICD PCR引物,以小鼠胎脑cDNA文库为模板,利用PCR技术扩增DLL1-ICD cDNA。将扩增出的基因片段克隆至pEGFP-C1载体,进行DNA序列测定。并进一步将测序正确的pEGFP-C1-DLL1-ICD真核表达载体转染HEK293细胞。结果显示:成功克隆了DLL1-ICD的cDNA片段,测序结果与基因文库中的DLL1-ICD序列一致,并在转染真核细胞后获得了较好地表达。表明pEGFP-C1-DLL1-ICD真核表达载体构建成功,并在293细胞中成功表达了DLL1-ICD-EGFP融合蛋白。研究结果为进一步探讨Notch信号通路与神经系统疾病的分子机制奠定基础。

Construction of Mammalian Expression Vector of DLL1-ICD and Transfection

To construct and identify mammalian expression vector of DLL-ICD(Delta-like1 inter cell domain) using cloning and transfection technique,the pair of primers were decigned based on musculus cDNA sequence,and the DLL1-ICD was amplified by PCR.And then the PCR product was cloned into the vector pEGFP-C1 which was analyzed by sequence.Finally,mammalian expression vector of pEGFP-C1-Dll1-ICD was transfected into 293 cells.Musculus Dll1-ICD cDNA was cloned and analyzed,and the sequence of gene from pEGFP-C1-Dll1-...