Abstract:

Objective   To study the type-specific regulation promoter in neural stem cells. Methods   The mouse nestin promoter sequence (4000bp), core sequence (400bp) and intron-2 were amplified with PCR. With pcDNA3.1 as a template, CMV, CMV + intron-2, nestin promoter sequence, nestin promoter sequence + intron-2, nestin promoter core sequence, and nestin promoter core sequence +intron-2 were constructed and used as promoters to regulate expression of enhanced green fluorecence protein(EGFP). These six kinds of recombinant plasmids were transfected into nestin+ and nestin-cells. Then EGFP expression was observed under a fluorescence microscope and determined by flow cytometry. Results   Both nestin promoter sequence and nestin promoter core sequence, with strong regulatory capacity, could non-specifically regulate expression of the EGFP gene in nestin+ and nestin- cells. Once  constructed with intron-2, they could only regulate expression of the EGFP gene in nestin+ cells. The CMV promoter sequence had non-specific regulatory capacity after being fused with intron-2. Conclusion   Nestin promoter sequences and intron-2 could collaboratively, typespecifically regulate expression of foreign genes in nestin+ cells.

Key words: Gene, nestin; Promoter regions(Genetics); Genes, regulator