The Construction and Identification of Eukaryotic Expression Vector pEGFP-N1-hTERT

[Objective] The aim of this study is to construct eukaryotic expression vector pEGFP-N1-hTERT and observe its expression in eukaryotic cells.[Method] The eukaryotic expression vector pEGFP-N1-hTERT was constructed with pC1-neo-hTERT and pEGFP-N1 plasmids,and the accuracy of human telomerase reverse transcriptase(hTERT) gene fragment was confirmed by double enzyme digestion and DNA sequencing analysis.After transfecting the pEGFP-N1-hTERT eukaryotic expression vectors into rat fetal neural stem cells(NSCs),the protein localization of human telomerase reverse transcriptase were indirectly observed through green fluorescent protein in the cells,and the correctness of constructed pEGFP-N1-hTERT was certificated by RT-PCR and Western Blot analysis.[Result] The structure of pEGFP-N1-hTERT eukaryotic expression vector was correct and it could express in eukaryotic cells.[Conclusion] This study laid a foundation for the establishment of immortalized NSCs line in rats.