Recombination fusion protein Trx-TAT-hMsrA and application thereof to aspect of nerve cell protection

A trx-tat-hmsra and fusion protein technology, applied in the field of genetic engineering, can solve the problems of poor druggability, difficult to penetrate the blood-brain barrier, difficult to enter cells, etc.

Inactive Publication Date: 2011-02-16
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is difficult for protein-based biotechnology drugs to enter cells, and it is also difficult for peripheral drug delivery to cross the blood-brain barrier
In order to overcome these shortcomings of po...

Method used

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  • Recombination fusion protein Trx-TAT-hMsrA and application thereof to aspect of nerve cell protection
  • Recombination fusion protein Trx-TAT-hMsrA and application thereof to aspect of nerve cell protection
  • Recombination fusion protein Trx-TAT-hMsrA and application thereof to aspect of nerve cell protection

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Experimental program
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Embodiment 1

[0051] Embodiment 1 prepares fusion protein of the present invention

[0052] 1. Construction of recombinant Trx-TAT-hMsrA fusion protein expression vector

[0053] The constructed vector map is attached figure 1 .

[0054] Detailed steps of vector construction:

[0055] (1). Sequencing: The Pcmv-XL4 vector containing the hMsrA gene (Beijing Yixing Industry Technology Co., Ltd.) was sent to Beijing Aoke Biotechnology Co., Ltd. for sequencing. The sequencing results were consistent with the human-derived MsrA gene in the gene bank Consistent, indicating that the plasmid is available.

[0056] (2). Design primers: In order to extract the hMsrA gene from the Pcmv-XL4 vector and switch it to the pet32a vector, design a pair of primers with BamHI and XhoI as restriction sites, and add Upper Tat sequence, 5' end sequence: 5-CGCGGATCCGGTTACGGTCGTAAGAAACGTAGACAGCGCAGACGTGGTCGCCCTCCCATGCTCTCGGC-3;

[0057] 3' end sequence:

[0058] 5-CCGCTCGAGTTATTTTTTAATACCCACTGGGCAGGACACGC-3, ...

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Abstract

The invention discloses a preparation method of a fusion protein Trx-TAT-hMsrA and application thereof to an aspect of nerve cell protection. The fusion protein Trx-TAT-hMsrA comprises a TAT protein transduction structure domain, thioredoxin Trx and anthropogenic methionine sulfoxide reductase hMsrA. The fusion protein of the invention has various biological effects for reducing the oxidization of functional protein methionine residues, treating oxidative stress injury, relieving the relevant nerve cell pathological change of parkinsonism, preventing and treating calcium ion overloading and the like, simultaneously has good capability for entering nerve cells through the epicyte, and belongs to novel biotechnological medicine candidates with good druggability.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a fusion protein (recombinant Trx-TAT-hMsrA fusion protein) formed by fusing TAT protein transduction domain, human methionine sulfoxide reductase hMsrA and thioredoxin Trx ), and its preparation method and application in nerve cell protection. Background technique [0002] A large number of modern neuroscience studies have shown that pathological damage mediated by oxygen free radicals is involved in the occurrence and development of a variety of major diseases of the nervous system. The brain's rich lipid content and relative lack of antioxidant enzyme levels make it more vulnerable to oxidative stress. Various factors (such as abnormally folded protein accumulation, ischemia-reperfusion injury, etc.) induce neurons to produce more reactive oxygen species (ROS) through pathways such as mitochondrial aerobic respiration, such as O 2- ., H 2 o 2 , OH an...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K47/48A61P25/00A61K38/44A61K38/17A61P25/28A61P9/10C12N15/62A61P25/16A61K47/42
Inventor 陈建国曾建华吴鹏飞王芳张醉杨远坚
Owner HUAZHONG UNIV OF SCI & TECH
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