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Method for clturing transgenic fowls

A technology for transgenic and poultry, applied in the field of preparing transgenic poultry, can solve the problems that the chimera method is not a transgenic chicken, the reverse transcription method is difficult to be accepted by the public, and the microinjection method is technically difficult.

Inactive Publication Date: 2006-04-19
INST OF LIVESTOCKS C A A S +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] It can be seen from the above that the microinjection method is technically difficult and has a low success rate; the reverse transcription method is difficult to be accepted by the public; the sperm carrier method has not been completely successful;

Method used

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  • Method for clturing transgenic fowls
  • Method for clturing transgenic fowls
  • Method for clturing transgenic fowls

Examples

Experimental program
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Effect test

Embodiment 1

[0025] In this example, the expression plasmid pOVΔβ was used, and the transgenic test was carried out according to the method of the present invention. Wherein, the expression plasmid pOVΔβ is the commercially available plasmid pCMVβ( figure 1 , Promega company product) after HindIII and SalI digestion and then self-ligated into pCMVβΔ, pCMVβΔ was then cut with PstI and SmaI to open a gap, and T4 ligase was used to insert the pre-prepared chicken ovalbumin promoter (OV ), the specific construction process of the plasmid is detailed in Chinese patent application 99122092.7.

[0026] The specific steps for preparing transgenic poultry are as follows:

[0027] Dissolve 50 micrograms of pOVΔβ plasmid DNA in 200 microliters of sterile distilled water, slowly add to 200 microliters (equivalent to 200 micrograms) of liposomes (products from Boehringer Mannheim, product number 1781995), mix well, and incubate at room temperature 45 minutes spare;

[0028] Collect 3 milliliters of ...

Embodiment 2

[0037] In this example, the expression plasmid pOVβ was used, and the transgenic test was carried out according to the method of the present invention. Wherein, the expression plasmid pOVβ is obtained by digesting pOVΔβ with BamHI, removing the SV40 fragment, and then inserting the 646bp fragment at the 3' end of the chicken ovalbumin gene prepared in advance. The specific construction process of the plasmid is detailed in Chinese patent application 99122092.7.

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Abstract

A method for preparing transgenic fowls including chicken, duck, goose, etc includes embedding foreign DNA (or gene) in hollow spiral liposome, incubating along with sperm to fuse sperm with liposome, electric shock for increasing the penetrability of sperm membrane to allow DNA to come in sperm easily, and artificial fertilization.

Description

technical field [0001] The invention relates to a method for preparing transgenic poultry, especially transgenic chicken. Background technique [0002] Transgenic animals are a high-tech biotechnology that has only been developed in recent years. Through this technology, foreign genes can be introduced into the genome of animals, so that the transgenic animals have new characteristics. Such as rapid growth, disease resistance, and stress resistance, so as to achieve the purpose of cultivating new excellent varieties; or introducing foreign genes to change the structure of genes in the body, so that certain genetic diseases in animals and humans can be treated. Or cultivate animals that express foreign proteins in animals, that is, "animal bioreactors". To achieve the above purpose, the preparation method of transgenic animals is the primary problem. [0003] Compared with mammalian embryos, chicken eggs are surrounded by yolk and located in the center of the blastodisc. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/027C12N15/87
Inventor 李建凡梁志清王小珂
Owner INST OF LIVESTOCKS C A A S
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