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Method for constructing chicken model for transgenosis and production exogenous protein

An exogenous protein and transgenic technology, applied in recombinant DNA technology, animal husbandry, etc., can solve the problems of transient expression of transgenic protein, inflammatory response, etc., and achieve high efficiency, low introduction dose, and good repeatability.

Inactive Publication Date: 2009-01-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the host itself does not have these proteins, it will naturally recognize them as non-self antigens and eliminate them, resulting in the transient expression of transgenic proteins and even triggering an inflammatory response

Method used

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  • Method for constructing chicken model for transgenosis and production exogenous protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1. Introduction of exogenous protein-bovine serum albumin (BSA) to eggs of different embryonic ages

[0019] Take 30 white Laihang chicken eggs (purchased from China Agricultural University Breeder Farm) with embryo ages of 3, 4, 5, 6 and 7 days respectively. The egg determines the position of the embryo, and then uses a pencil to mark the edge of the air chamber on the eggshell on the opposite side of the embryo, so that the embryonic vascular network can be effectively avoided when punching, and the mechanical effect of the embryonic blood vessels when introducing foreign proteins can be avoided. damage. Make a punch mark 0.3-0.5cm below the marked edge of the air chamber. In order to maintain the air pressure balance between the air chamber and the embryo chamber, make another punch mark on the top of the air chamber, wipe the punch mark with alcohol cotton, and use The dissecting needle quickly pierced two small holes (1mm in diameter) at the egg punch mark...

Embodiment 2

[0023] Example 2. Detection of immune tolerance results of transgenic model chickens inoculated with bovine serum albumin at different embryonic ages by ELISA

[0024] After bovine serum albumin was emulsified with Freund's adjuvant (Freund's complete adjuvant was used for antigen emulsification during the first immunization, and Freund's incomplete adjuvant was used for antigen emulsification at other age stages), it was obtained in Example 1 above. On the 20th, 30th, 40th and 50th days after hatching, transgenic model chickens inoculated with bovine serum albumin of different embryo ages were treated with 100 μl of bovine serum albumin solution with a concentration of 1 g / L for 4 days. After the first subcutaneous injection, on the 7th day after the fourth subcutaneous injection, the serum of the transgenic model chickens was taken for ELISA detection. At the same time, the chicken serum from untreated chicks in the embryonic stage after being immunized four times at the abo...

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Abstract

The invention discloses a method for constructing a chick model which is used for transgene and generating heterologous protein. The method of the invention comprises the following steps: 1) the heterologous protein is introduced to the yolk sac of a breeding egg; 2) the breeding egg in step 1) is incubated still for 46 to 50 hours and incubated continuously until hatching birds, thus obtaining the chick model which is used for transgene and generating the heterologous protein. The method of the invention has the advantages of simple and easily obtained operation condition, low cost, high maneuverability, low introducing dosage of the heterologous protein, good repeatability and high hatching rate. Besides, the method of the invention has little influence on chick embryo development, the hatching rate can reach 73 percent to 83 percent, the durable time of immune tolerance is relatively long, and the chick model which is used for transgene of 60 days still has immune tolerance to specific heterologous protein antigens.

Description

technical field [0001] The present invention relates to a method for constructing a transgenic model chicken for producing exogenous protein. Background technique [0002] The establishment of DNA recombination technology in the 1970s enables heterologous DNA to be introduced into eukaryotic cells. This gene transfer technology provides humans with the ability to genetically alter higher animal cells, and has become an important tool in the preparation of transgenic animals and Fundamentals of gene therapy. Using DNA recombination technology, people have successfully obtained a variety of transgenic animals including mice, cattle, pigs, rabbits, chickens, fish, nematodes and sea urchins. In 1990, transgenic methods were also used in human gene therapy research. . For many higher animals, especially vertebrates with a well-established immune system, the gene delivery system has potential immunostimulatory properties, and the expression of heterologous genes can easily cause...

Claims

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Application Information

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IPC IPC(8): A01K67/02A01K67/027C12N15/09
Inventor 李赞东赵晨
Owner CHINA AGRI UNIV
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