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Polymorphism of first exon in ADAMTSI gene

An exon and gene technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, sugar derivatives, etc., can solve the problem of no exon polymorphism of ADAMTS1 gene

Inactive Publication Date: 2004-08-11
CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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AI Technical Summary

Problems solved by technology

[0003] Before this application, there was no related report on the polymorphism of exon 1 of ADAMTS1 gene

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Gene extraction and sequencing

[0026] DNA was extracted from human blood by the conventional phenol-chloroform method, and the concentration was corrected to 20ng / ul for conventional PCR amplification. Primers are:

[0027] Sense primer: 5'-ctggcgcact gcttctactc-3' (SEQ ID NO: 3)

[0028] Antisense primer: 5'-tgaagtcgcg tgggatagat a 3' (SEQ ID NO: 4)

[0029] The amplified 445bp product was purified and subjected to DNA sequencing.

Embodiment 2

[0031] SNP acquisition

[0032] Direct sequencing of the ADAMTS1 gene. The measured sequences of each sample are compared to obtain sequence differences and obtain SNPs.

Embodiment 3

[0034] Detection kit

[0035] Prepare a detection kit for detecting ADAMTS1 gene-related disease susceptibility, which contains the following primer pairs that can amplify 322 SNPs:

[0036] Sense primer: 5'-ctggcgcact gcttctactc-3' (SEQ ID NO: 3)

[0037] Antisense primer: 5'-tgaagtcgcg tgggatagat a-3' (SEQ ID NO: 4)

[0038] Chromatographic analysis of the amplified product and the normal control is carried out with a denaturing high-performance liquid chromatograph (DHPLC), and the G->C type SNP at position 322 can be easily detected.

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Abstract

A single nucleotide polymorphism (SNP) for the extron No.1 of ADAMTS1 gene and a method for analyzing the SNP and activity of said extron No.1 are disclosed. Said SNP is the G->C polymorphism at site No.322 in its sequence shown by SEQ ID No.1 and can cause the change Ala(A)->Pro(P) at the site No.108 of coding protein, so changing protein activity.

Description

technical field [0001] The invention relates to the single nucleotide polymorphism of the first exon of ADAMTS1 gene. The present invention also relates to a method for analyzing the allelic mutation of the first exon of ADAMTS1 gene. Background technique [0002] ADAMTS1 is a secreted protein with a signal peptide at the N-terminus, a zinc-binding site, and a cysteine-rich region. The ADAMTS family of proteins all function in proteolytic processes. Some researchers have found that both in vivo and in vitro results show that ADAMTS1 is extremely effective in destroying the angiogenesis process, and the effect is more prominent than that of Thrombospondin-1, endostatin and ADAMTS8. (J. Biol. Chem. 274:23349-23357, 1999.). [0003] Before this application, there was no relevant report on the polymorphism of exon 1 of ADAMTS1 gene. Contents of the invention [0004] The purpose of the present invention is to provide the polymorphism of the first exon of ADAMTS1 gene and i...

Claims

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Application Information

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IPC IPC(8): C07H21/00C12N15/11C12P19/34C12Q1/68
Inventor 黄薇施锦绣奚慧峰金力
Owner CHINESE NAT HUMAN GENOME CENT AT SHANGHAI
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