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CYP2D6 gene fragment comprising 74G>A mutation, coded protein fragment and applications thereof

A technology of fragments and nucleic acid fragments, applied in the field of biology, can solve the problems of drug toxicity and side effects, insufficient treatment, and differences in drug efficacy.

Active Publication Date: 2014-01-22
BEIJING HOSPITAL
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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 CYP2D6 gene is the main reason for the significant difference in CYP2D6 enzyme activity among individuals, and individuals carrying different CYP2D6 genotypes can cause great differences in drug efficacy, and even serious Drug toxicity or inadequate treatment of

Method used

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  • CYP2D6 gene fragment comprising 74G>A mutation, coded protein fragment and applications thereof
  • CYP2D6 gene fragment comprising 74G>A mutation, coded protein fragment and applications thereof
  • CYP2D6 gene fragment comprising 74G>A mutation, coded protein fragment and applications thereof

Examples

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Embodiment 1

[0069] Example 1: Identification of new mutation sites in human CYP2D6 gene

[0070] In this example, blood samples were collected from normal healthy people of the Han nationality, genomic DNA in the blood was extracted, and sequencing primers were designed to amplify and sequence the 9 exons of the CYP2D6 gene to analyze whether there were mutation sites in the CYP2D6 gene.

[0071] 1) DNA extraction:

[0072] Take a 5ml venous EDTA anticoagulated blood sample from the subject; then follow the common salting out method and / or use a special DNA extraction kit (purchased from Omega, USA) to extract the genomic DNA of the blood sample to be tested.

[0073] 2) PCR amplification:

[0074] Amplification primers were designed to amplify the 9 exon 5 sequences of the CYP2D6 gene in the obtained genomic DNA samples. The sequences of the amplification primer pairs are shown in Table 1.

[0075] Use 30μL PCR reaction system, including: 1×GC PCR buffer, 1.5mM MgCl 2 , 100ng of geno...

Embodiment 2

[0089] Example 2: Analysis of enzyme metabolic activity in vitro

[0090] According to the existing research results, the metabolic activity of the wild type (*1 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 *10 type is higher than that of the *10 type. Type 2 is lower (see literature 6, 7). 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 (for example, the metabolic activity of the enzyme expressed by the genotype can be inferred). activity compared to the metabolic activity of the wild-...

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Abstract

The invention belongs to the fields of biology, and relates to a mutation of the single base in the 74th site of a CYP2D6 allelic gene, wherein the site is mutated from G to A. In specific, the invention relates to a nucleic acid fragment comprising the mutation site, a corresponding coded protein fragment thereof, a reagent for identifying the mutation site, a detecting method, and applications of the site, particularly applications of identification of the site for guiding pharmacy.

Description

technical field [0001] The invention belongs to the field of biology and relates to the single base mutation of the 74th position of CYP2D6 allele. More specifically, the present invention relates to a nucleic acid fragment containing the mutation site and its corresponding encoded protein fragment, a reagent for identifying the mutation site, a detection method and the application of identifying the site in guiding medication. Background technique [0002] Cytochrome P450 2D6 (CYP2D6) is one of the important members of the CYP enzyme family. Its gene is located on chromosome 22, including 9 exons, with a total length of 9432bp (GenBank registration number M33388, and the exon region is located at positions 1620-5909). The amount of cytochrome in the human body only accounts for 2% to 4% of the total amount of liver enzymes, but it participates in the metabolism of 20% to 30% of clinical drugs, such drugs include antidepressants, antiarrhythmic drugs, antipsychotic drugs, s...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/11C12N9/02C12Q1/68
Inventor 蔡剑平胡国新戴大鹏耿培武蔡杰
Owner BEIJING HOSPITAL
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