Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Human TSHR extracellular fusion protein and preparation method thereof

A fusion protein, extracellular segment technology, applied in biochemical equipment and methods, chemical instruments and methods, microorganism-based methods, etc., can solve problems such as low TSH binding rate

Inactive Publication Date: 2010-12-29
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Transfection of hTSHR gene into insect cells by baculovirus vector can obtain milligram quantities of TSHR extramembrane protein, but most of these proteins are insoluble in water and have low binding rate with TSH

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human TSHR extracellular fusion protein and preparation method thereof
  • Human TSHR extracellular fusion protein and preparation method thereof
  • Human TSHR extracellular fusion protein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0047] The present invention provides a recombinant protein capable of efficiently expressing TSHR antigenicity. By establishing Pichia pastoris engineering bacteria that can secrete and express human TSHR, a highly expressed secreted protein with TSHR antigenicity can be obtained in its good in vitro expression system. Soluble recombinant protein. The present invention will be described in detail below, which is an explanation of the present invention rather than a limitation.

[0048] 1. Construction of the expression vector pPICZαA-hTSHRecd:

[0049] a. Cloning of hTSHRecd extracellular segment gene

[0050] According to the human TSHR gene sequence (NM_000369.2) in GenBank, a pair of oligonucleotide-specific primers for amplifying hTSHRecd cDNA coding region were designed and synthesized:

[0051] Upstream primer P1: cc ctcgag aa aagagggtgt tcgtctccac 30;

[0052] Downstream primer P2: gct ctagag c atcatcatca tcttttctca ggaacttgta g 41;

[0053] P1 contains the Xho...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses human TSHR extracellular fusion protein and a preparation method thereof. The N end of the human TSHR extracellular fusion protein is connected with exogenous secretion signal peptide, and the C end thereof is connected with a purification marker sequence; when the fusion protein is expressed and purified, the fusion protein the N end of which is connected with the exogenous secretion signal peptide is resected, and the specific amino acid sequence is shown in SEQ ID No.1. The invention obtains high-expression soluble recombinant protein with the antigenic secretion form of TSH receptor in a good in-vitro expression system by constructing recombinant pichia pastoris containing pPICZ alpha A-hTSHRecd expression vector.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the recombination and bioengineering preparation of human TSH receptors, in particular to a human TSHR extracellular fusion protein and a preparation method thereof. Background technique [0002] Autoimmune thyroid disease (AITD) is an organ-specific disease, including Graves' disease (GD), Hashimoto's thyroiditis (Hashimoto's thyroiditis, HT) and idiopathic myxedema (Primary myxedema, PM), its incidence rate accounts for the first place in endocrine system diseases. Thyrotropin receptor (TSHR) is the main antigen that induces AITD. [0003] The molecular weight of human TSHR is 84.5KD, and it reaches 100KD after glycosylation, which is closely related to the pathogenesis of AITD. Human TSHR mainly exists on the thyroid cell membrane and belongs to one of the members of the guanine nucleotide regulatory protein (G protein) coupled receptor superfamily; it consists of three parts: the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/81C12P21/02C12R1/84
Inventor 施秉银田竹芳杨琪伍丽萍徐利高雷
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com