CYP2C9 gene segment comprising 394C>T, 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 drug side effects, insufficient treatment, and differences in drug efficacy

Active Publication Date: 2013-04-17
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 394C>T, coded protein segment and application thereof
  • CYP2C9 gene segment comprising 394C>T, coded protein segment and application thereof
  • CYP2C9 gene segment comprising 394C>T, coded protein segment and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0065] Example 1: Identification of new mutation sites of 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) Extract DNA:

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

[0069] 2) PCR amplification:

[0070] Design amplification primers to amplify the 9 exon sequences of the CYP2C9 gene in the obtained genomic DNA sample. The sequence of the amplification primer pair is shown in Table 1.

[0071] Using 50μl PCR reaction system, including: 1×PCR buffer, 1.5mM MgCl 2 , 100-150ng genomic DNA, 0.2μM upstream and downstream primers, 0.4mM ...

Embodiment 2

[0085] Example 2: Analysis of enzyme metabolic activity

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

[0087] In this...

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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 394th site of SEQ ID NO.2 corresponding to a CYP2C9 gene, wherein the mutation site is mutated into T from wild type C. 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 method for detecting the mutation site.

Description

Technical field [0001] The invention belongs to the field of biology and relates to single base mutations. More specifically, the present invention relates to a mutation site of the CYP2C9 gene relative to the 1001th position of SEQ ID NO. 1 or the 394th position of SEQ ID NO. 2, a nucleic acid fragment containing the mutation site and the corresponding protein fragment encoded thereby . The invention also relates to reagents and detection methods for identifying the mutation site, and the application of identifying the 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 amount of human liver microsomal CYP enzymes. About 10-16% of clinically commonly used drugs are oxidized and metabolized by CYP2C9, which mainly include tolbutamide, S-warfarin, phenytoin, glipizide, glibenclamide, tolazimide, losartan, and erucal Besartan and many non-s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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