Method for constructing chicken model for transgenosis and production exogenous protein

An exogenous protein, transgenic technology, applied in recombinant DNA technology, animal husbandry and other directions, can solve the problems of inflammatory response, transient expression of transgenic protein, etc., achieve the effect of low introduction dose, high efficiency acquisition, and solve the problem of individual immune response.

Inactive Publication Date: 2010-11-03
CHINA AGRI UNIV
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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

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Effect test

Embodiment 1

[0018] Embodiment 1, introducing exogenous protein-bovine serum albumin (BSA) to the breeding eggs of different embryonic ages

[0019] 30 eggs (purchased from the breeder farm of China Agricultural University) with 3, 4, 5, 6, and 7 days of embryonic age were taken, and the surface of the eggshell was wiped with alcohol cotton for disinfection, and then irradiated by a microscopic light source. Egg to determine the position of the embryo, and then use a pencil to mark the edge of the air cell on the eggshell on the opposite side of the embryo, so that the embryonic vascular network can be effectively avoided when punching holes, and the mechanical damage to the embryonic blood vessels can be avoided when foreign proteins are introduced. damage. Make a punching mark 0.3-0.5cm below the edge of the marked air chamber. In order to maintain the air pressure balance between the air chamber and the germ chamber, make another punching mark on the top of the air chamber, wipe the pun...

Embodiment 2

[0023] Embodiment 2, Utilize enzyme-linked immunoassay (ELISA) to detect the immune tolerance result of the transgenic model chicken inoculated with bovine serum albumin at different embryonic ages

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

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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 invention relates to a method for constructing a transgenic model chicken for producing foreign protein. Background technique [0002] The establishment of DNA recombination technology in the 1970s allowed heterologous DNA to be introduced into eukaryotic cells. This gene transfer technology provided humans with the ability to genetically change higher animal cells, and became a tool for preparing transgenic animals and carrying out The basis of gene therapy. Using DNA recombination technology, people have successfully obtained a variety of transgenic animals including mice, cows, pigs, rabbits, chickens, fish, nematodes and sea urchins. In 1990, transgenic methods were also used in the research of human gene therapy . For many higher animals, especially vertebrates with a complete immune system, the gene delivery system is potentially immunostimulatory, and the expression of heterologous genes can easily cause the host to generate an immune resp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/02A01K67/027C12N15/09
Inventor 李赞东赵晨
Owner CHINA AGRI UNIV
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