Method for authenticating copy number of target genes in transgenic animal

A technology for transgenic animals and target genes, applied in the field of identification of target gene copy number in transgenic animals, can solve time-consuming and labor-intensive problems and achieve consistent results

Inactive Publication Date: 2010-11-24
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
View PDF0 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Facts have proved that it is indeed an effective and accurate method, but it is not difficult to find in the actual operation process

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for authenticating copy number of target genes in transgenic animal
  • Method for authenticating copy number of target genes in transgenic animal
  • Method for authenticating copy number of target genes in transgenic animal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Determining the copy number of exogenous genes in transgenic animals

[0029] 1. Preparation of transgenic animals (GH transgenic pigs)

[0030] 1. Using TRE as a framework, construct a porcine GH gene vector (TRE-GH58 vector) (see figure 1 ).

[0031] Synthesize a DNA fragment containing TRE promoter, multiple cloning site and SV40polyA (named TRE-MSC-SV40polyA gene, its nucleotide sequence is shown in sequence 6 in the sequence table, synthesized by Beijing Aoke Biotechnology Co., Ltd.). The DNA fragments synthesized above were immediately placed in ice water at 95°C for 10 minutes, and after treatment, they were double-enzymatically digested with ZraI. (purchased from Promega, Cat. No. FD3219) after ZraI. and PciI double digestion and ligation with the aforementioned TRE-MSC-SV40 polyA fragment (ligase was purchased from Promega, Cat. Quanshijin Company, Cat. No. CD201) Competent Escherichia coli; the bacterial solution was coated on an agarose gel plat...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for authenticating the copy number of target genes in a transgenic animal. The method provided by the invention comprises the following steps of: taking genome DNA of the transgenic animal as a template; performing fluorescence quantitative PCR by respectively using a target gene primer and a reference gene primer to respectively obtain a fluorescence quantitative PCR result of the target genes and reference genes of the transgenic animal; taking a standard substance with the target genes in a different copy number as the template; performing the fluorescence quantitative PCR by using the target gene primer and the reference gene primer respectively to obtain the fluorescence quantitative PCR result of the target genes and the reference genes in the standard substance; establishing a standard curve relative to the difference values of the cycle indexes of the target genes and the reference genes and the natural logarithm of the copy number of the target genes; and determining the copy number of the target genes in the transgenic animal. The method has the characteristics of simplicity, easy implementation, and accuracy and reliability because the results are consistent by using a conventional Southern blot method.

Description

technical field [0001] The invention relates to a method for identifying the copy number of a target gene in a transgenic animal. Background technique [0002] Transgenic animals (transgenic animals) use laboratory methods to introduce the target genes that people need into their genomes, so that the foreign genes can be integrated with the genes of the animals themselves, and proliferate with cell division, and can be expressed in animals. An animal that is stably passed on to offspring. The foreign structural genes integrated into the animal genome are called transgenes, and the proteins encoded by the transgenes are called transgenic products, which affect animal traits through transgenic products. If the transgene is passed on to offspring, a line or population of transgenic animals is formed. With the continuous deepening of research on transgenic animals and the continuous improvement of experimental techniques, transgenic technology has been more widely used. Almost...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68G01N21/64
Inventor 李奎鞠辉明白立景崔文涛牟玉莲唐中林杨述林程文科王志伟
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap