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Method for constructing esophageal epithelial tissue p53 specific knockout mouse esophageal precancerous lesion model

A technology of epithelial tissue and construction methods, applied in chemical instruments and methods, biochemical equipment and methods, recombinant DNA technology, etc., can solve problems that are not involved, can not be treated and studied, and achieve the effect of long survival period

Inactive Publication Date: 2018-10-30
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A model of tissue canceration can be established by knocking out p53, but this canceration model is a general canceration model, and it cannot conduct targeted research on the cancerous process and post-cancerous treatment of specific tissues
Currently, specific cancers targeting esophageal epithelial tissue have not been involved in existing reports

Method used

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  • Method for constructing esophageal epithelial tissue p53 specific knockout mouse esophageal precancerous lesion model
  • Method for constructing esophageal epithelial tissue p53 specific knockout mouse esophageal precancerous lesion model
  • Method for constructing esophageal epithelial tissue p53 specific knockout mouse esophageal precancerous lesion model

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Experimental program
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Embodiment 1

[0039] The steps for constructing a p53 knockout mouse esophageal epithelial tissue-specific carcinogenesis model are described as follows:

[0040] 1. Hybrid pairing

[0041] 1. Screen the p53 homozygous mice and Cre-positive mice of the F0 generation mice;

[0042] ED-L2-Cre mice are heterozygous positive, and negative mice will appear during the breeding process, so positive mice need to be screened, namely ED-L2-Cre + / - The mice used in the experiment; and the introduced p53floxed mice are also heterozygous positive (p53 flox / flox and p53 flox / wild ), it needs preliminary identification and screening, and the selected genotype is B6.p53 flox / floxThe mice were used for the experiment, so ED-L2-Cre + / - mice and B6.p53 flox / flox The mice are F0 generation mice.

[0043] 2. F0 generation mice were crossed to obtain F1 generation mice

[0044] B6.p53 flox / flox Female / male mice with ED-L2-Cre + / - The male / female mice in the same cage were paired and crossed with each ot...

Embodiment 2

[0084] The steps for constructing a p53 knockout mouse esophageal epithelial tissue-specific carcinogenesis model are described as follows:

[0085] 1. Hybrid pairing

[0086] 1. Screen the p53 homozygous mice and Cre-positive mice of the F0 generation mice;

[0087] From the identification and screening of ED-L2-Cre mice and p53floxed mice, the genotype was selected as ED-L2-Cre + / - mice and B6.p53 flox / flox The mice are F0 generation mice.

[0088] 2. F0 generation mice were crossed to obtain F1 generation mice

[0089] B6.p53 flox / flox Female / male mice with ED-L2-Cre + / - Male / female mice were paired and crossed in cages; after the F1 generation mice were weaned (about 21 days after birth), the male and female pups were separated into cages, ear-marked, and 0.5-1cm of the mouse tail was cut off, and the tissue was lysed and extracted. DNA, PCR identification of the genotype of F1 generation mouse DNA, lysing tissue and extracting DNA, PCR identification method is the s...

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Abstract

The invention particularly relates to a method for constructing an esophageal epithelial tissue p53 specific knockout mouse esophageal precancerous lesion model. The method comprises the following steps: 1) performing multi-generation hybridization on B6.p53flox / flox mice and ED-L2-Cre+ / -mice, identifying mouse genotypes and screening to obtain p53 knockout mice, wherein p53 knockout mouse genotypes are B6.p53flox / flox and ED-L2-Cre+ / -; 2) inducing the p53 knockout mice by using a chemical carcinogen to form the p53 knockout mouse epithelial tissue specific carcinogenesis model. The mouse esophageal precancerous lesion model is successfully constructed by utilizing the method.

Description

technical field [0001] The invention belongs to the technical field of animal model construction methods, and in particular relates to a construction method of an esophagus precancerous lesion model in p53-specific knockout mice of esophageal epithelial tissue. Background technique [0002] Methylbenzylnitrosamine is an asymmetric nitrosamine, which is an alkylating agent of DNA, and can be catabolized into benzaldehyde and methane diazohydroxide under the action of cytochrome P450 oxidase, thereby causing DNA Addition 0 6 - Methylguanine is formed, which causes a mismatch with thymine during DNA replication, resulting in the transformation of DNA from G:C pairing to A:T pairing, resulting in genetic material damage; in addition, 06-methylguanine can be combined with DNA sequences Cross-linking of opposite cytosine residues in the DNA prevents DNA from replicating. Studies have shown that the enzymes required for NMBzA metabolism are specifically present in esophageal epit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/0331C07K14/47C12N15/8509
Inventor 赵继敏刘康栋陈新焕许岩岩董子明江亚南张晓艳杨婉景路静朱艳艳朱历历
Owner ZHENGZHOU UNIV
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