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CYP2D6 gene segment containing 1693A>G mutation, coded protein fragment thereby 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 treatment, differences in drug efficacy, and insufficiency

Active Publication Date: 2014-03-05
蔡剑平
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  • Summary
  • 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 segment containing 1693A>G mutation, coded protein fragment thereby and applications thereof
  • CYP2D6 gene segment containing 1693A>G mutation, coded protein fragment thereby and applications thereof
  • CYP2D6 gene segment containing 1693A>G mutation, coded protein fragment thereby and applications thereof

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Embodiment 1

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

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

[0071] 1) Extract DNA:

[0072] Take 5ml of intravenous EDTA anticoagulated blood sample from the 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).

[0073] 2) PCR amplification:

[0074] Design amplification primers to amplify the 9 exon sequences of CYP2D6 gene in the obtained genomic DNA sample. The sequence of the amplification primer pair is shown in Table 1.

[0075] Using 30μL PCR reaction system, including: 1×GC PCR buffer, 1.5mM MgCl 2 , 100ng of genomic DNA, 0....

Embodiment 2

[0088] Example 2: In vitro enzyme metabolic activity analysis

[0089] According to the existing research results, the metabolic activity of wild type (*1 type) is relatively high for various drugs, while the metabolic activity of *2 type is significantly lower than that of wild type, and the metabolic activity ratio of *10 type* Type 2 is lower (see literature 7, 8). 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).

[0090] In th...

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Abstract

The invention belongs to the biological field, and relates to single base mutation of the 1693th locus of CYP2D6 allele. The locus is mutated into G from A. The invention concretely relates to a nucleic acid segment containing the mutation locus, a corresponding coded protein fragment, agents for identification of the mutation locus, a detection method and applications of the locus, especially applications of the identification of the locus in medication guidance.

Description

technical field [0001] The invention belongs to the field of biology and relates to the single base mutation of the 1693rd 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,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/11C12Q1/68C12N9/02
Inventor 蔡剑平胡国新戴大鹏耿培武蔡杰
Owner 蔡剑平
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