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Human ICAM-1 extramembranous I-III domain gene expression product ,its prep. and application in radioimmunological technology

An adhesion molecule and expression product technology, applied in the field of genetic engineering, can solve the problems of no thyroid extract and high price, and achieve the effects of easy purification, simple operation and low cost

Inactive Publication Date: 2003-10-15
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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Problems solved by technology

[0009] At present, there is no thyroid extract of ICAM-1 at home and abroad, and ICAM-1 genetic engineering products obtained through eukaryotic systems abroad are expensive

Method used

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  • Human ICAM-1 extramembranous I-III domain gene expression product ,its prep. and application in radioimmunological technology
  • Human ICAM-1 extramembranous I-III domain gene expression product ,its prep. and application in radioimmunological technology
  • Human ICAM-1 extramembranous I-III domain gene expression product ,its prep. and application in radioimmunological technology

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Experimental program
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Embodiment Construction

[0034] (1) Acquisition of human ICAM-1 extramembrane I-III functional region genes and their identification:

[0035] TRIzol method was used to extract total RNA from the thyroid tissue of patients with Graves disease after surgery, and RT-PCR method was used to synthesize and amplify the human ICAM-1 extramembrane I-III functional region gene, using 100bp DNA gradient standard as the nucleic acid molecular weight standard, 1% agarose The result of gel electrophoresis to identify the PCR product showed a clear electrophoresis band at about 868bp, which was consistent with the size of the target gene fragment to be amplified. figure 1 , in the figure 1 Bgl I digestion product, 2 100bp DNALadder.

[0036] Bgl I digestion of PCR products, 1% agarose gel electrophoresis to identify the results of digestion products, showing that two clear electrophoresis bands can be seen at 505 and 363bp, which is consistent with the theoretical calculation results. figure 2 , in the figure 1 ...

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Abstract

The present invention relates to gene engineering technology. The I-III functional encoding gene outside intercellular adhesion molecule-1 (ICAM-1) membrane is recombined to expression plasmid pET42a, transferred into prokaryotic expression bacteria, and purified to obtain expression product GST-ICAM-1. The expression product is used as antigen in immunizing rabbit to obtain rabbit anti-human polyclonal antibody. Thus prepared expression product of human I-III functional encoding gene outside ICAM-1 membrane in prokaryotic system is easy to purify, low in cost and high in yield. The human serum sICAM-1 radioimmunoassay technology is developed and used in clinical test and has important significance in diagnosing the treating effect of Graves' disease.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a human intercellular adhesion molecule-1 extramembrane I-III functional region gene expression product, a preparation method thereof and an application of the product in radioimmunoassay technology. Background technique [0002] Autoimmune thyroid disease is a frequently-occurring disease, including Graves' disease (GD), chronic lymphatic thyroiditis (HT), etc., and its pathogenesis has not yet been fully understood. Recent studies have shown that intercellular adhesion molecule-1 (ICAM-1) is closely related to its pathogenesis. Intercellular adhesion molecules refer to a group of glycoprotein substances expressed on the cell surface that have intercellular adhesion. They are involved in mutual recognition, mutual adhesion, signal transmission and transfer, immune cell mutual recognition and Interaction, recirculation and migration of leukocytes and other functions....

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/395C07K14/705C07K16/28C12N15/12C12N15/63C12N15/70
Inventor 方佩华吕枚张志友何红鹏高硕
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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