TAT-PAP fusion protein as well as preparation method and application thereof

A technology of fusion proteins and protein transduction domains, which can be applied in the fields of peptide/protein components, chemical instruments and methods, hybrid peptides, etc., can solve problems affecting drug research and development, and achieve low-cost effects

Inactive Publication Date: 2014-12-17
XIAN MEDICAL UNIV
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  • Abstract
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Problems solved by technology

Protein transduction technology is very important in neuroscience research, because the main regulatory substances of cells and intercellular physiological reactions in the central nervous system are poly...

Method used

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  • TAT-PAP fusion protein as well as preparation method and application thereof
  • TAT-PAP fusion protein as well as preparation method and application thereof
  • TAT-PAP fusion protein as well as preparation method and application thereof

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Embodiment Construction

[0029] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0030] The TAT-PAP fusion protein of the present invention is a fusion protein TAT-PAP formed from the protein transduction domain PTD of the transcriptional transactivator TAT of human type I immunodeficiency virus HIV-I and the prostatic acid phosphatase PAP.

[0031] The fusion protein has the amino acid sequence shown in SEQ ID NO:1.

[0032] The TAT-PAP fusion protein of the present invention fuses the PTD of TAT derived from human immunodeficiency virus HIV-1 with PAP to construct pTAT-PAP recombinant expression plasmid vector, and obtains TAT-PAP with biological activity through expression and purification The fusion protein has the advantages of low cost and high activity.

[0033] The TAT-PAP fusion protein of the present invention has the characteristic that TAT can carry out protein transmission, breaks through the limitation that t...

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Abstract

The invention relates to a TAT-PAP fusion protein. The TAT-PAP fusion protein is formed by protein transduction domain PTD of a trans-activator of transcription of human I type immunodeficiency virus HIV-I and prostatic acid phosphatase PAP. A preparation method of the TAT-PAP fusion protein comprises the following steps: selecting a pTAT-HA expression carrier containing Nco I and Xho I restriction enzyme cutting site sequences at the two ends respectively; synthesizing a pUC57-PAP carrier; carrying out double digestion on the pTAT-HA expression carrier and the pUC57-PAP carrier, and recycling respectively; obtaining a pTAT-PAP recombinant expression plasmid carrier; transforming the pTAT-PAP recombinant expression plasmid carrier into escherichia coli DE3 competent cells; and inducing expression of the fusion protein. The TAT-PAP fusion protein has the advantages that not only is the original penetrating power of TAT remained, but also activity of a foreign protein PAP is hardly influenced, so that the TAT-PAP fusion protein has a broad prospect in clinical application of a macromolecule protein medicine on diabetic neuropathic pain.

Description

technical field [0001] The invention belongs to the technical field of protein transduction, and specifically relates to a TAT-PAP fusion protein, and also relates to a preparation method and application of the TAT-PAP fusion protein. Background technique [0002] Diabetic neuropathic pain is one of the most serious complications of diabetes. It is caused by extensive degeneration and inflammatory demyelination of peripheral sensory nerves in the state of chronic diabetes. , abnormal pain (allodynia) and hyperalgesia (hyperalgesia), so that the limbs function abnormally, thus causing serious harm to the patient's body and mind. At present, for the clinical treatment of diabetic neuropathic pain, in addition to strictly controlling blood sugar and improving microcirculation according to the etiology, some classic analgesic drugs, such as morphine and non-steroidal anti-inflammatory drugs, etc., are very effective in relieving diabetic pain. There is evidence that tricyclic a...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61P25/02A61K47/48A61K38/46C12N15/70
Inventor 崔媛媛苟兴春方科徐浩张妮李妮赵湘辉
Owner XIAN MEDICAL UNIV
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