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Method of detecting mononucleotide pleomorphism of CYP2D6 gene ninth exon

A single nucleotide polymorphism and detection method technology, which is applied in the field of detection of single nucleotide polymorphisms in exon 9 of the CYP2D6 gene, can solve problems such as undiscovered and achieve significant theoretical effects

Inactive Publication Date: 2008-01-23
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

[0003] According to the search of the existing literature, there is no report related to the SNP (single nucleotide polymorphism) of the ninth exon (G4046A) of the CYP2D6 gene of the present invention

Method used

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Embodiment Construction

[0015] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific condition in the following examples, usually according to routine conditions, such as people such as Sambrook, molecular cloning: the condition described in the laboratory handbook (New York: Cold Spring Harbor Laboratory Press, 1989), or according to manufacture conditions recommended by the manufacturer.

[0016] step 1

[0017] Isolation and sequencing of a nucleic acid

[0018] Primer design:

[0019] Using Primer 5.0 software, a pair of allele-specific nucleic acid primers were designed using the GenBank database CYP2D6 gene (AY545216) exon 9 and the exon-intron junction sequence as templates, which were synthesized by Invitrogen.

[0020] Primer information:

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Abstract

The invention relates to a testing method of CYP2D6 gene exon 9's single nucleotide polymorphism, and meanwhile relates to a separation nucleic acid and an allel-specific nucleic acid primer. The method comprises steps as described below: firstly the confirmation of the 1332 nucleic acid showed in the SEQ ID No: 1 in the human CYP2D6 gene exon 9, then test of the existence of the single nucleotide polymorphism, specifically a separation nucleic acid with the SEQ ID NO: 1 and the 1322 position is A, an allel-specific nucleic acid primer with the length of 15 to 50bp and specifically hybridizes and amplifies the amplified products of the 1322 nucleic acid polymorphism showed in the SEQ ID No: 1 in the human CYP2D6 gene exon 9. The invention can be used to research the relation between CYP2D6 gene polymorphism in Chinese people and the clinical drug safety, and provide guidance to the development of new drugs.

Description

technical field [0001] The present invention relates to nucleic acid and its primers and detection method in the field of gene technology, in particular to an isolated nucleic acid, an allele-specific nucleic acid primer and a single nucleotide polynucleotide (SNP) of the ninth exon of CYP2D6 gene state-of-the-art detection method. Background technique [0002] Cytochrome P450 2D6 (CYP2D6), also known as isoquinidine hydroxylase, is a member of cytochrome P450. CYP2D is the first cytochrome oxidase P450 found to have genetic polymorphisms, and at least 21 CYP2Ds have been found in mammals. Human CYP2D is located at 22q13.1 and is composed of CYP2D6, CYP2D7P and CYP2D8P. CYP2D7P and CYP2D8P are pseudogenes, and only CYP2D6 has functional protein expression. CYP2D6 is mainly expressed in the liver, and CYP2D6 has a total of 497 amino acids. CYP2D6 not only oxidatively metabolizes certain endogenous steroid hormones, but also is the main metabolizing enzyme for 20-25% of com...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/25C12N15/11G01N33/02C07H21/00
Inventor 贺林秦胜营唐吉敏沈陆
Owner SHANGHAI JIAO TONG UNIV
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