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Fusion protein

A technology of fusion protein and binding protein, applied in the field of fusion protein, can solve the difficult problems of phosphatidylserine eversion tracking and drug targeting

Inactive Publication Date: 2016-11-30
南通睿科医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above methods are difficult to track and drug-target the externalization of phosphatidylserine in vivo.

Method used

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Embodiment 1

[0040] Example 1 Membrane Expression Functional Annexin A5 Fusion Protein Construction and Expression

[0041] This embodiment provides a fusion protein, its construction method is as follows figure 1 As shown in B, it includes the first peptide segment and the second peptide segment. The first peptide segment includes the secretion signal peptide and the phosphatidylserine binding protein peptide segment, and the second peptide segment segment includes the connecting peptide segment and the transmembrane peptide segment. , and signal transduction peptides with cell regulation functions. The specific experimental steps are as follows:

[0042] 1. Construction of the second peptide region of the fusion protein

[0043] The commercial eukaryotic expression plasmid pEGFP-N1 (SEQ ID NO: 1) was selected as the vector. Gene synthesis encodes the nucleotide sequence of the second peptide region of the fusion protein (SEQ ID NO: 2), which sequence includes the Hind III site, the Ko...

Embodiment 2

[0051] Example 2 Membrane expression of non-functional annexin A5 fusion protein construction and expression

[0052] This embodiment provides a fusion protein, its construction method is as follows figure 1 As shown in A, it includes the first peptide region and the second peptide region. The first peptide region includes the secretion signal peptide and the phosphatidylserine binding protein peptide, and the second peptide region includes the connecting peptide and the transmembrane peptide. part. The specific experimental steps are as follows:

[0053] 1. Construction of the second peptide region of the fusion protein

[0054] Gene synthesis encodes the nucleotide sequence of the second peptide region of the fusion protein (SEQ ID NO: 9), which sequence includes Hind III site, Kozak sequence gccacc, translation initiation codon ATG, and 3xGGGGS peptide chain coding sequence from the 5' end Nucleic acid sequence, CD8a fragment (382nt-629nt in P01732-1), BamHI site.

[00...

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Abstract

The invention relates to the field of biomedicine, particularly to a fusion protein. The fusion peptide comprises a first peptide fragment region and a second peptide fragment region, the C terminal of the first peptide fragment region is connected to the N terminal of the second peptide fragment region, wherein the first peptide fragment region includes a secretion signal peptide, and a phosphatidylserine binding protein peptide segment, the second peptide fragment region includes a connecting peptide segment and a transmembrane peptide segment, the phosphatidylserine binding protein peptide segment includes an phosphatidylserine antibody or a peptide segment in specific bonding with phosphatidylserine in annexin. The fusion protein provided by the invention has the characteristics of cell membrane protein and the ability of combining with a lipid bilayer structure, cytokine induced killers or stem cells expressing the protein can have the ability of tissue infiltration or enrichment at disease tissues and injured parts, and the fusion protein can be used for tracer, imaging, chemotherapy or phototherapy of disease tissues.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a fusion protein. Background technique [0002] Phosphatidylserine (PS), a ubiquitous phospholipid, is usually located in the inner layer of eukaryotic cell membranes and is one of the cell membrane components. Under normal physiological conditions, there is asymmetry in the distribution of plasma membrane lipids, which is due to the existence of a phospholipid translocase in the plasma membrane, which can specifically transport phosphatidylserine (PS) and phosphatidylethanolamine (PE) to At the same time, another flippase in the plasma membrane is responsible for transporting phosphatidylserine (PS) and phosphatidylcholine (PC) to the outer leaflet of the plasma membrane. Both enzymes act on the membrane simultaneously, but flippase reacts 10 times slower than translocase. These two enzymatic reactions are carried out independently, that is, when the activity of flippase is inhibite...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/85
CPCC07K16/18C07K7/08C07K14/47C07K2319/02C07K2319/03C12N15/85C12N2800/107
Inventor 郝晓勇黄招琴
Owner 南通睿科医药科技有限公司
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