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.

Inactive Publication Date: 2014-07-02
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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Problems solved by technology

Both increase the chance of recurrence
Moreover, no matter whether it is corticosteroids, immunoglobulins or plasma exchange, these drugs or treatments are not cell-specific, and have the disad...

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  • Recombinant vector for expressing light-sensitive type adenylate cyclase, applications and construction method of the recombinant vector and treating system for demyelination
  • Recombinant vector for expressing light-sensitive type adenylate cyclase, applications and construction method of the recombinant vector and treating system for demyelination
  • Recombinant vector for expressing light-sensitive type adenylate cyclase, applications and construction method of the recombinant vector and treating system for demyelination

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Embodiment 1

[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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Abstract

The invention relates to a recombinant vector used for expressing light-sensitive type adenylate cyclase, applications and a construction method of the recombinant vector, and a treating system for demyelination. The recombinant vector comprises a viral vector, and a shuttle plasmid wrapped in the viral vector and carrying the gene sequence of the light-sensitive type adenylate cyclase. The gene sequence of the light-sensitive type adenylate cyclase comprises specific promoters of an oligodendroglia cell and a Schwann cell. The recombinant vector is adopted to infect a demyelination part so that the gene sequence of the light-sensitive type adenylate cyclase is only expressed in the oligodendroglia cell and the Schwann cell. Under stimulation of light, the cAMP level in the two cells can be adjusted directionally, thus promoting remyelination and stable myelin sheath growth and achieving an objective of treatment of the demyelination by cell specificity.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a recombinant vector expressing light-sensitive adenylate cyclase, its application, construction method and a treatment system for demyelinating diseases. Background technique [0002] Demyelinating diseases are neurological diseases in which demyelination is the main or initial lesion. It can occur in the central nervous system or peripheral nervous system. The myelin sheath of the central nervous system is composed of oligodendrocytes, while the myelin sheath of the peripheral nervous system is composed of Schwann cells. Demyelinating diseases are divided into two types: myelin destruction and myelin formation disorders. Demyelinating diseases are one of the insurmountable challenges facing clinical medicine, especially neurology today. In recent years, the number of cases with myelin damage has increased significantly. [0003] At present, there is no specific and ef...

Claims

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Application Information

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IPC IPC(8): C12N15/86C12N15/861C12N15/867A61K48/00A61P25/00
Inventor 谢维娇王丹冯晓娜秋云海
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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