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Method and kit for detecting mouse genotype knockout of apolipoprotein M gene

A gene knockout mouse, apolipoprotein technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as unclear mechanism of action, and achieve high sensitivity, simple operation, and easy judgment.

Inactive Publication Date: 2015-06-10
THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have shown that insulin, leptin, estrogen, hyperglycemia and many cytokines can regulate the expression of apolipoprotein M, and apolipoprotein M may also be related to the occurrence and development of diseases such as obesity, diabetes and fetal growth and development, but Its mechanism of action is unclear

Method used

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  • Method and kit for detecting mouse genotype knockout of apolipoprotein M gene
  • Method and kit for detecting mouse genotype knockout of apolipoprotein M gene
  • Method and kit for detecting mouse genotype knockout of apolipoprotein M gene

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Embodiment Construction

[0030] 1. Materials and methods.

[0031] 1.1. Specimen source: The nucleic acid fragment from exon 2 to exon 5 of the C57BL / 6 mouse apolipoprotein M gene was replaced with the exogenous carrier sequence Lox-Neo-LoxP by gene knockout technology, Apolipoprotein M gene knockout heterozygous mice (hereinafter referred to as knockout heterozygous mice) that can be stably inherited have been successfully obtained, and apolipoprotein M gene knockout homozygous mice can be obtained through breeding ( Hereinafter referred to as knockout homozygous mice).

[0032] After breeding and breeding, knockout heterozygous mice and knockout homozygous mice can grow and reproduce normally, and their coat color, eating, activity, body temperature and quality are similar to those of wild homozygous mice (without gene knockout). type mice showed no significant difference.

[0033] 1.2. Main reagents and instruments: Ezup column animal genome DNA extraction kit [Sangon Bioengineering (Shanghai) Co...

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PUM

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Abstract

The invention discloses a method and kit for detecting mouse genotype knockout of an apolipoprotein M gene. The method comprises the following steps: designing and synthesizing primers and probes; extracting DNA (Deoxyribonucleic Acid) of a mouse; and carrying out real-time fluorescent quantitative PCR (Polymerase Chain Reaction) detection. The kit comprises a nucleic acid extraction reagent, a PCR reaction reagent, RNase-free water, the primers and the probes. The designed primers comprise a wild homozygotic type forward primer, a wild homozygotic type reverse primer, a wild homozygotic type probe, a knockout homozygotic type forward primer, a knockout homozygotic type reverse primer and a knockout homozygotic type probe. The method disclosed by the invention can be used for completing genotype detection within 1 hour and is relatively low in cost and simple in operation. The method disclosed by the invention is accurate in result and easy in judgment and also has relatively high sensitivity.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and a kit for detecting the genotype of apolipoprotein M gene knockout mice. Background technique [0002] Gene knockout technology is through homologous recombination between exogenous DNA and chromosomal DNA to carry out precise fixed-point modification and genetic modification. It has the characteristics of strong specificity and stable inheritance of chromosomal DNA and target fragments. An ideal experimental method for modifying biological genetic material. Gene knockout mice use gene knockout technology to inactivate the target gene at a specific site in the genome of normal experimental mice. Ideal experimental materials for target genes. Gene knockout technology has been greatly developed from vector construction to cell screening to establishment of animal models. [0003] Apolipoprotein M (Apolipoprotein M, ApoM) was discovered by Xu and Dahlback in ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2561/113C12Q2563/107
Inventor 罗光华郑璐于洋
Owner THE FIRST PEOPLES HOSPITAL OF CHANGZHOU
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