Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for breeding genetically modified animal by using recombinant adenovirus vector

A technology for recombinant adenovirus and transgenic animals, applied in the fields of molecular biology, genetic engineering, and cell biology, can solve the problems of low overall efficiency, simple and efficient transgenic methods, early embryo damage, etc.

Inactive Publication Date: 2009-09-02
NORTHWEST A & F UNIV
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has unavoidable defects: it is necessary to use a microscopic operating system to precisely inject exogenous DNA into the male pronucleus of fertilized eggs, which will inevitably cause damage to early embryos, and the integration efficiency of exogenous genes is random. overall low efficiency
To sum up, at present, there is no simple and efficient transgenic method in the world

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 breeding genetically modified animal by using recombinant adenovirus vector
  • Method for breeding genetically modified animal by using recombinant adenovirus vector
  • Method for breeding genetically modified animal by using recombinant adenovirus vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation method of human thrombin III transgenic animal

[0021] (1) Synthesis, cloning and sequence determination of target protein (such as human thrombin III) cDNA:

[0022] 1. Extraction of total RNA from human liver tissue

[0023] (1) Cut a small piece of frozen tissue, quickly weigh it, and then transfer it into the freshly prepared lysis buffer, and add 1 ml of lysis buffer for every 50 mg of tissue.

[0024] (2) Homogenize the liver tissue to break the cells, pre-filter and centrifuge once, and collect the filtrate.

[0025] (3) Add an equal volume of chloroform, shake and mix, centrifuge and aspirate the supernatant.

[0026] (4) Add an equal volume of 70% ethanol, mix gently, and centrifuge to obtain total RNA.

[0027] (5) Dissolve the precipitate in Rnase-free water treated with 0.01% DEPC, and perform content determination by conventional methods.

[0028] 2. Synthesis of total cDNA by reverse transcription

[0029] (1) Take 1~2μg of total breast ...

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 relates to a method for breeding a genetically modified animal by using a recombinant adenovirus vector. The invention adopts a technical proposal that: a replication-defective adenoviral framework plasmid vector carries a foreign target gene sequence to produce a recombinant adenovirus vector in a packaging cell (293, 911 and other cells) containing a sequence of an Ela region; the recombinant adenovirus vector is combined with a donor egg or fertilized egg cell; and the donor egg or fertilized egg cell is first cultured and developed in vitro and then transferred into the body of a receptor animal for normally breeding so as to produce the genetically modified animal which is genetically stable and has functions of different target genes. The method for breeding the genetically modified animal is high in success rate. The genetically modified animal can be used in the confirmation of functional genes, the expression of pharmaceutical proteins (including organs, blood and mammary glands), and the building of models of genetically modified animal having functions of different target genes and animal genetic diseases.

Description

Technical field [0001] The invention belongs to the technical fields of cell biology, molecular biology and genetic engineering, in particular to a method for preparing transgenic animals by using adenovirus as a carrier. Background technique [0002] With the advent of the post-genomic era of life sciences, the research and application of genes and their products has attracted more and more attention from researchers. Transgenic animal technology is one of the most effective technical methods. At present, internationally The strategy is to process fertilized eggs, eggs and sperm to obtain transgenic animals from three aspects. In view of the instability of egg and sperm processing methods, in the preparation of transgenic animals, the fertilized egg processing method is still the most effective and classic method. Now, the existing method of preparing transgenic animals is processed for fertilized eggs. The summary is as follows. [0003] Prokaryotic phase microinjection of fore...

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): C12N15/861A01K67/027C12R1/93
Inventor 李青旺韩增胜赵红卫胡建宏宁国柱
Owner NORTHWEST A & F UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com