Method for regulating and controlling nerve stem cell differentiation, and restraining differentiation
A neural stem cell, silencing technology, applied in the field of inhibiting the differentiation of neural stem cells, can solve problems such as lack of cure methods
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[0038] This embodiment includes the following steps:
[0039] 1. Construction of recombinant fluorescent plasmid vector pEGFP-C1-DCF1
[0040] pEGFP-C1 is a mammalian expression vector, the green fluorescent protein GFP encoded by the reporter gene can emit green fluorescence at 510nm when excited by near-ultraviolet light at 395nm or blue light at 470-490nm. The upstream and downstream primers were designed according to the full-length gene sequence of DCF1, and the full-length gene DCF1 was amplified by PCR using the neural stem cell mRNA library as a template. The DCF1 gene was inserted into the multiple cloning site of the fluorescent plasmid vector pEGFP-C1 to construct the recombinant vector pEGFP-C1-DCF1.
[0041] The sequences of the upstream and downstream primers are as follows. In order to facilitate the connection with the vector, the upstream primer of the Kpn1 adapter site is put in: 5'-CTAT GGTACC ATGGCGGCGCCAAAGGGGAA-3'
[0042] Kpn1
[00...
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