Human MCHR1 epitope domain gene expression-associated protein MCHR1-C and encoding gene thereof

A technology of MCHR1-C and antigenic epitopes, which is applied in receptors/cell surface antigens/cell surface determinants, animal/human proteins, genetic engineering, etc., can solve problems such as difficult treatment and unclear pathogenesis

Active Publication Date: 2011-01-26
许爱娥 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the unclear pathogenesis of the disease, it is difficult to treat

Method used

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  • Human MCHR1 epitope domain gene expression-associated protein MCHR1-C and encoding gene thereof
  • Human MCHR1 epitope domain gene expression-associated protein MCHR1-C and encoding gene thereof
  • Human MCHR1 epitope domain gene expression-associated protein MCHR1-C and encoding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Case Collection

[0045] From January 1, 2008 to December 30, 2009, the serum of 100 patients with vitiligo who were not combined with other diseases and had not been treated with immunosuppressants in the past 3 months and had obvious progress in the dermatology clinic of Hangzhou Third People's Hospital were collected. Among them, there were 82 cases of common type (10 cases of generalized, 8 cases of localized, 16 cases of acral, 33 cases of sporadic, 15 cases of sporadic combined with acral), 18 cases of segmental type; 48 cases of male, There were 52 females, aged 16-67 years, with an average age of 32 years, and the course of disease ranged from 1 month to 20 years, with an average of 3.5 years. Serum samples from 30 normal people, including 16 males and 14 females, aged 18 to 47 years, with an average age of 31, were collected from healthy individuals. All collected cases were signed by patients with informed consent and approved by the Medical Committe...

Embodiment 2

[0046] Example 2: Synthesis of Human MCHR1 Epitope Region Gene MCHR1-C

[0047] According to the literature reports, human MCHR1 antigen epitope regions 51-80, 85-98, 154-158 and 254-260 were artificially synthesized according to the nucleotide sequence of the full-length human MCHR1 gene (Genbank accession number: NM005297.3), A BamH I restriction site was introduced at the 5' end of the gene, an Xho I restriction site and a stop codon sequence were introduced at the 3' end, and the size of the synthetic gene MCHR1-C was 183bp. This work was completed by Nanjing GenScript Biotechnology Co., Ltd. The measured sequence of the synthesized antigen epitope region gene is consistent with SEQ ID NO:2.

Embodiment 3

[0048] Example 3: Construction of human MCHR1 epitope region gene MCHR1-C expression vector

[0049] The amplified MCHR1-C fragment was purified, digested with BamHI / xhoI (purchased from Treasure Bioengineering (Dalian) Co., Ltd.) and connected to the prokaryotic expression vector pGEX-4T-2 (gifted by Professor Zhang Chuanxi, Institute of Insect Science, Zhejiang University ) BamH I / Xho I site, T4DNA ligase (purchased from Treasure Bioengineering (Dalian) Co., Ltd.) after connection, the recombinant plasmid was transformed into BL21 Escherichia coli competent cells, and the recombinant transformant was screened, and the screened recombinant plasmid was After pGEX-MCHR1-C was digested and identified by PCR, it was sent to Shanghai Sangong Company for sequencing. The measured sequence of the inserted antigenic epitope fragment is consistent with SEQ ID NO:2.

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Abstract

The invention provides a human melanin concentrating hormone receptor-1 (MCHR1) epitope domain gene expression-associated protein MCHR1-C, an encoding gene and a recombinant vector thereof and an application of the polypeptide in identifying antoantibody in serum of a vitiligo patient. The human MCHR1 epitope domain gene expression-associated protein MCHR1-C of the invention has an amino acid sequence shown in SEQ ID No:1. The human MCHR1 epitope domain gene expression-associated protein MCHR1-C of the invention has the beneficial effects that an polypeptide capable of effectively identifying the antoantibody in the serum of the vitiligo patient is provided; and the application of the human MCHR1 epitope domain gene expression-associated protein and the polypeptide in detecting MCHR1 antibody of pigmentary dermatosis, in particular the autoantibody in the serum of the vitiligo patient, thereby laying foundation for further new drug screening.

Description

(1) Technical field [0001] The invention relates to a human melanin concentrating hormone receptor-1 (Melanin Concentrating Hormone Receptor 1, MCHR1) antigen epitope region gene expression-related protein MCHR1-C, its coding gene, a recombinant vector, and the use of this polypeptide in identifying vitiligo Application of autoantibodies in patient serum. (2) Background technology [0002] Vitiligo is a skin pigmentation loss disease caused by the destruction of melanocytes. Vitiligo patients are spread all over the world, accounting for about 1-2% of the world's population. If calculated as 1% in my country, there are more than 10 million patients with vitiligo. The damage caused by the disease to beauty has seriously affected the patients' mental health, normal social activities and employment choices, and reduced the quality of life of patients. Suicide due to the disease. At present, because the pathogenesis of the disease is not yet clear, the treatment is difficult. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705C12N15/12C12N15/63C12Q1/68G01N33/53
Inventor 许爱娥关翠萍周妙妮
Owner 许爱娥
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