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CYP2C9 gene segment comprising 287G>C, coded protein segment and application thereof

A technology of fragments and nucleic acid fragments, applied in the field of biology, can solve problems such as differences in drug efficacy, insufficiency, drug toxicity and side effects, etc.

Active Publication Date: 2013-04-10
蔡剑平
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] According to current clinical research, this polymorphism of CYP2C9 gene is the main reason for the great difference in CYP2C9 enzyme activity among individuals, so it can cause great differences in drug efficacy among individuals carrying different CYP2C9 genotypes, Even severe drug side effects or inadequate treatment

Method used

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  • CYP2C9 gene segment comprising 287G>C, coded protein segment and application thereof
  • CYP2C9 gene segment comprising 287G>C, coded protein segment and application thereof
  • CYP2C9 gene segment comprising 287G>C, coded protein segment and application thereof

Examples

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Embodiment 1

[0065] Example 1: Identification of new mutation sites in human CYP2C9 gene

[0066] In this example, 2127 blood samples were collected, genomic DNA in the blood was extracted, sequencing primers were designed to amplify and sequence the 9 exons of the CYP2C9 gene, and screen the mutation sites of the CYP2C9 gene

[0067] 1) DNA extraction:

[0068] Take 5ml of venous EDTA anticoagulated blood sample from each subject; then extract the genomic DNA of the blood sample to be tested according to the common salting-out method and / or using a special DNA extraction kit (purchased from Omega, USA). .

[0069] 2) PCR amplification:

[0070] Amplification primers were designed to amplify the 9 exon sequences of the CYP2C9 gene in the obtained genomic DNA sample. The sequences of the amplification primer pairs are shown in Table 1.

[0071]Use 50μl PCR reaction system, including: 1×PCR buffer, 1.5mM MgCl 2 , 100-150ng of genomic DNA, both upstream and downstream primers are 0.2μM, ...

Embodiment 2

[0085] Example 2: Analysis of Enzyme Metabolic Activity

[0086] According to the existing research results, the metabolic activity of the wild type to various drugs is relatively high, while the metabolic activity of the *2 type is significantly lower than that of the wild type, and the metabolic activity of the *3 type is lower than that of the *2 type ( See refs 18, 19, 21, 22). Therefore, there is a consensus in the field that the metabolic activity of enzymes expressed by the same genotype on specific substrates can represent the metabolic activity on other substrate drugs. Thereby, according to the specific substrate metabolic activity data of the enzyme expressed by a certain genotype, the metabolic activity of the enzyme expressed by the genotype to other substrate drugs can be deduced (such as, the metabolic activity of the enzyme expressed by the genotype can be inferred). The activity was compared with the metabolic activity of the enzyme expressed by the wild type...

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Abstract

The invention belongs to the field of biology, and relates to single base mutation. More particularly, the invention relates to a mutation site of a 287th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into C from wild type G. The invention also relates to a nucleic acid segment, a coded protein segment and application thereof. The invention also provides an allele-specific oligonucleotide, kit and detection method for detecting the mutation site.

Description

technical field [0001] The invention belongs to the field of biology and relates to single base mutation. More specifically, the present invention relates to the mutation site of CYP2C9 gene relative to the 1001st position of SEQ ID NO.1 or the 287th position of SEQ ID NO.2, the nucleic acid fragment comprising the mutation site and the corresponding encoded protein fragment thereof . The invention also relates to a reagent and a detection method for identifying the mutation site, and the application of the identification site in guiding medication. Background technique [0002] CYP2C9 is the most important member of the CYP2C subfamily of the cytochrome P450 enzyme family, accounting for about 20% of the total CYP enzymes in human liver microsomes. About 10-16% of commonly used clinical drugs are oxidatively metabolized by CYP2C9, mainly including tolbutamide, S-warfarin, phenytoin, glipizide, glibenclamide, toratimide, losartan, urethane Drugs such as besartan and many ...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/11C12N9/02C12Q1/68
Inventor 蔡剑平戴大鹏徐仁爱胡国新杨丽萍
Owner 蔡剑平
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