Investigation of neural stem cell-specific regulatory promoter elements
- Authors:
- Honghua Yuan
- Ankang Hu
- Li Zhang
- Xiaorong Zhu
View Affiliations
Affiliations: Department of Neurobiology, Xuzhou Medical College, Xuzhou, Jiangsu 221002, P.R. China, Laboratory Animal Center, Xuzhou Medical College, Xuzhou, Jiangsu 221002, P.R. China
- Published online on: June 18, 2012 https://doi.org/10.3892/etm.2012.614
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Pages:
405-408
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Abstract
The present study aimed to investigate neural stem cell (NSC)-specific regulatory promoter elements. PCR was employed to amplify the full sequence (4,000 bp) and core sequence (400 bp) of the promoter and intron-2 of the mouse nestin gene. pcDNA3.1 was used as a template to construct 6 different recombinant plasmids. CMV, CMV + intron-2, the full sequence of the nestin gene promoter, the full sequence of the nestin gene promoter + intron-2, the core sequence of the nestin gene promoter and the core sequence of the nestin gene promoter + intron-2 were independently used as promoters to regulate EGFP expression. The 6 recombinant plasmids were independently used to transfect nestin-positive and nestin-negative cells, and the expression of EGFP was observed under a fluorescence microscope. At the same time, flow cytometry was carried out to measure the proportion of cells positive for EGFP. The results showed that the full sequence and core sequence of the nestin gene promoter non-specifically regulated EGFP expression in cells and exhibit potent regulatory potency. The full sequence or core sequence of the nestin gene promoter which was fused with intron-2 can only regulate the EGFP expression in nestin-positive cells. CMV + intron-2 have non-specific regulation of EGFP alone. Thus, we conclude that the full sequence of the nestin gene promoter which is fused with intron-2 can specifically regulate the expression of exogenous genes in nestin-positive cells.
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