Recombinant vector for expressing light-sensitive type adenylate cyclase, applications and construction method of the recombinant vector and treating system for demyelination
An adenylate cyclase and demyelinating disease technology, which is applied in the field of treatment systems for demyelinating diseases, and can solve problems such as the inability to change the progression and course of the disease, the difficulty in achieving remyelination, and the lack of cell specificity.
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[0071] Construction of Recombinant Vector for Expressing Light-sensitive Adenylyl Cyclase
[0072] (1) The MBP promoter was used as the specific promoter of oligodendrocytes and Schwann cells. After PCR amplification of the MBP promoter, restriction endonuclease digestion and T4 DNA ligase ligation were used to amplify the expanded The MBP promoter was inserted into the plasmid with EYFP (such as Figure 4Shown), get MBP promoter recombination data, verify its expression, such as Figure 5 As shown, the specific expression of the target protein is normal.
[0073] (2) Artificially synthesized light-sensitive adenylate cyclase (bPAC) gene sequence, and inserted the bPAC gene sequence downstream of the MBP promoter. The sequencing was successful, and the light-sensitive adenylyl cyclase gene sequence containing the MBP promoter was obtained (such as Figure 6 shown), denoted as MBP-bPAC. MBP-bPAC was inserted into the shuttle plasmid to obtain a shuttle plasmid carrying a li...
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