Investigation of neural stem cell-specific regulatory promoter elements

Yuan,Honghua; Hu,Ankang; Zhang,Li; Zhu,Xiaorong

doi:10.3892/etm.2012.614

September 2012 Volume 4 Issue 3

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September 2012 Volume 4 Issue 3

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Article

Investigation of neural stem cell-specific regulatory promoter elements

Authors:
- Honghua Yuan
- Ankang Hu
- Li Zhang
- Xiaorong Zhu
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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
Pages: 405-408
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Published online on: June 18, 2012

https://doi.org/10.3892/etm.2012.614
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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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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Investigation of neural stem cell-specific regulatory promoter elements

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