Yeast recombinant expression, purification and application of protein functional polypeptide with effect of preventing and treating atherosclerosis

A technology for atherosclerosis and protein function, applied in yeast recombinant expression, diagnosis, prevention and treatment of atherosclerosis, purification field

Active Publication Date: 2014-10-22
DALIAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Yeast recombinant expression, purification and application of protein functional polypeptide with effect of preventing and treating atherosclerosis
  • Yeast recombinant expression, purification and application of protein functional polypeptide with effect of preventing and treating atherosclerosis
  • Yeast recombinant expression, purification and application of protein functional polypeptide with effect of preventing and treating atherosclerosis

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Embodiment 1

[0068] Upstream primers:

[0069] 5'-CCG CTCGAGAAAAAGA AAAGCATCTTGTAAAGTACCTGTG-3' 39bp

[0070] Downstream primers:

[0071] 5'-GC TCTAGA TTATCA ATGATGATGATGATGATG GGAACCACCACCACC GCATGGCTTTACATCGGA-3' 65bp

[0072] The 5' end of the upstream primer contains a protective base, xho I restriction site, signal peptide recognition site and β 2 - Part of the amino acid coding gene sequence at the 5' end of the fifth functional domain of GPI; the 5' end of the downstream primer contains protective bases, Xbal I restriction site, stop codon, 6 His, GGGGS flexible peptide and β 2 -The complementary sequence of the amino acid coding gene at the 3' end of the fifth functional domain of GPI.

[0073] 2. The process of expression and purification of recombinant protein for prevention and treatment of atherosclerosis is as follows:

[0074] (1) β 2 - Obtaining the gene fragment of the fifth functional domain of GPI:

[0075] to contain β 2 - The T vector CTA727-1 of the...

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Abstract

The invention belongs to the field of biological engineering, and in particular to recombinant expression and purification of a fifth functional domain of an oxLDL binding protein human serum beta 2-glycoprotein I and the application thereof to diagnosis prevention and treatment of atherosclerosis. A designed primers pair is subjected to PCR amplification on a T vector of a beta 2-GPI gene fragment to obtain a gene fragment of a beta 2-GPI fifth functional domain; and pPICZ alpha A-DV is constructed by cloning, and is then subjected to power transformation to obtain a positive strain, which is subjected to recombination and purification to obtain a P.r beta 2-GPI-DV protein. The possibility of using the recombinant protein as a drug in the prevention and treatment of atherosclerosis and the feasibility of detecting the oxLDL content in serum are proved. The P.r beta 2-GPI-DV recombinant protein has purity above 90%, can reduce the content of oxLDL / beta 2-GPI binary compound in the serum, and then inhibit the uptake of oxLDL by scavenger receptors on the surface of macrophage, so as to achieve the effect of preventing and treating atherosclerosis; and because of the combination specificity with oxLDL, the P.r beta2-GPI-DV can realize the specific detection of oxLDL in serum.

Description

technical field [0001] The invention belongs to the field of bioengineering, in particular to oxLDL binding protein human serum beta 2 - Yeast recombinant expression and purification of the fifth functional domain of glycoprotein I and its application in the diagnosis, prevention and treatment of atherosclerosis. Background technique [0002] Atherosclerosis (AS) has become an important factor threatening human health with the improvement of people's living standards. The oxidation product of low density lipoprotein (LDL), oxidized low density lipoprotein (oxLDL) is considered to be the main inducing factor of atherosclerosis (C. K. Glass, and J. L. Witztum, Cell, 2001). oxLDL can interact with β in serum 2- Glycoprotein I (β 2 -glycoprotein I, β 2 -GP I) and form a stable binary complex, oxLDL / β 2 The -GPI binary complex can be used as an autoimmune antigen to form a ternary complex with autoantibodies in the serum of patients with systemic lupus erythematosus SLE or ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K14/435C07K1/36C07K1/22C07K1/34G01N33/68A61P9/10C12R1/19
Inventor 刘庆平李敬达迟彦王仁军
Owner DALIAN UNIV
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