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Recombinant galactose agglutinin-1 two-string protein and preparation method thereof

A galectin and protein technology, applied in the biological field, can solve problems such as tumor growth inhibition, and achieve the effect of simple separation and purification process and easy to expand production

Inactive Publication Date: 2012-05-23
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In Galectin-1-deficient mice, tumor growth was significantly inhibited due to the inhibition of angiogenesis

Method used

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  • Recombinant galactose agglutinin-1 two-string protein and preparation method thereof
  • Recombinant galactose agglutinin-1 two-string protein and preparation method thereof
  • Recombinant galactose agglutinin-1 two-string protein and preparation method thereof

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Embodiment Construction

[0045] The present invention provides biologically active recombinant Galectin-1 protein through biotechnology methods, and constructs prokaryotic expression vectors pET-22b(+)-Gal-1 and pET-22b(+)-Gal-1②, respectively transfecting BL21 construction projects Bacteria, and then induced expression by IPTG, chromatographic separation and purification after lysing to obtain recombinant Galectin-1 monomer protein or recombinant Galectin-1 binary protein with biological activity. The erythrocyte agglutination test confirmed that the two recombinant Galectin-1 proteins had obvious biological activities, and the biological activity of the recombinant Galectin-1 dimer protein was higher than that of the recombinant Galectin-1 monomeric protein. The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments, which are explanations of the present invention rather than limitations.

[0046] 1. Cloning of Galectin-1 monomer gen...

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Abstract

The invention discloses a recombinant galactose agglutinin-1 two-string protein and a preparation method thereof. The method specifically comprises the following steps of: providing a recombinant Galectin-1 protein with bioactivity through a biotechnological method; respectively transfecting BL21 to construct engineering bacteria through constructing prokaryotic expression vectors pET-22b(+)-Gal-1 and pET-22b(+)-Gal-1 (2); and then obtaining the recombinant Galectin-1 monomer protein or a recombinant Galectin-1 two-string protein through IPTG (Isopropyl Beta-D-1-Thiogalactopyranoside) induction expression and the chromatographic separation and passivation after dividing the bacteria. Through an erythrocyte agglutination test, it is proved that both the two recombinant Galectin-1 proteins have obvious bioactivity, and the bioactivity of the Galectin-1 two-string protein is higher than that of the Galectin-1 monomer protein.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the construction, prokaryotic expression, purification and identification of recombinant plasmids of target proteins, in particular to recombinant galectin-1 distring protein and its preparation method. Background technique [0002] 1. The basic characteristics of Galectin-1 [0003] Galectin-1 belongs to the Galectins family of endogenous lectins. So far, 14 members of the Galectins family have been found, at least 12 of which exist in the human body. All members of this family have carbohydrate recognition domains (Carbohydrate-recognition domains, CRD) with a structure of about 130 amino acids and have the ability to treat β-galactoside. Special affinity for two properties. Galectins can be divided into three types according to their molecular structures: ① Prototype, containing a single sugar recognition domain CRD, including Galectin-1, 2, 5, 7, 10, 13, 14; ② Chimeric type, cont...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12C12N15/70C07K1/18
Inventor 张英起李晶张伟
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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