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Technology for producing transgenic mammal through intrauterine exogenous gene injection method

A technology of transgenic animals and exogenous genes, applied in the field of preparing transgenic mammals, can solve the problems of SMGT technology doubts, and achieve the effect of avoiding in vitro culture and transplantation, and the method is simple and easy to operate.

Inactive Publication Date: 2015-11-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to Brinster RL et al. (1989) [34] Repeated experiments were done without positive results, and the SMGT technology was questioned

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] Embodiment 1: Preparation of transgenic mice

[0167] Ten sexually mature healthy female mice aged 6 weeks were taken. After the simultaneous estrus treatment, the female mice were reared in a cage with normal male mice of the same line at a ratio of 1:1, and the mating time was observed. After 48 hours of mating, pentobarbital sodium anesthesia was injected intraperitoneally at a dose of 0.0001 g per gram of body weight, and plasmid complexes were prepared while waiting for anesthesia. After the anesthesia reaction of convulsions appeared in the female rat, the female rat was placed in a prone position to protect her body. After entering the coma state, the back of the female mouse was removed symmetrically on both sides of the dorsal midline from about 5 cm below the head to expose the surgical site. Disinfect the skin, open the cortex and muscle layer respectively, pull out the white fat pad with ophthalmic tweezers, and the ovary is exposed, and use a 1ml syringe t...

Embodiment 2

[0169] Embodiment 2: the preparation of transgenic rabbit

[0170] 6 multiparous New Zealand female rabbits, superovulation treatment plan: 6 subcutaneous injections of FSH in the neck, each interval of 10-12 hours, the injection dose was reduced, the total dose was 60 IU, 12 hours after the last injection of FSH, hCG 100 was injected into the ear vein IU. Mate with the male rabbit after the last injection of hCG, and then mate again 12 hours later. 48 hours after the second mating, the female rabbit was anesthetized by Su Mianxin II and placed on a safety frame. A routine operation was performed along the abdominal linea alba at 5cm-7cm below the intersection of the last pair of nipples and the alba linea, using 1ml Inject 1ml of the exogenous gene pIRES2-EGFP with a working concentration of 0.5mg / ml-0.8mg / ml into the uterus with a syringe. After the injection, sprinkle an appropriate amount of penicillin and streptomycin mixture on the wound, suture the wound, and sprinkle ...

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PUM

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Abstract

A purpose of the present invention is to provide a method for producing transgenic mammal through an intrauterine exogenous gene injection method. The method is characterized in that an exogenous gene is injected into the maternal uterus within the specific period after the animal is subjected to breeding (the blastocyst formation stage), and parturition is normally performed so as to obtain the stably-genetic transgenic animal. Compared with other transgenic technologies in the prior art, the technology of the present invention has characteristics of simpleness, easy performing, low cost, and high success rate.

Description

technical field [0001] The invention relates to a method for preparing transgenic mammals. Background technique [0002] Mammalian transgenic technology is a new type of biotechnology that combines genetic engineering and embryo engineering. It is the introduction of artificially recombined foreign DNA into the genome of the recipient animal through artificial methods, or the removal of a segment of DNA in the recipient genome, thereby artificially changing the genetic information (genotype and phenotype) of the recipient animal , and a biotechnology in which this change can be passed on to future generations [1] . [0003] Chimeric mice since 1974 [2] Since its birth, mammalian transgenic technology has brought a new technological revolution to the disciplines of developmental biology, medicine and animal husbandry. With the development of molecular biology and biotechnology, mammalian transgenic technology has made great progress. With the help of transgenic animal mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89A01K67/027
Inventor 曾宪垠宋天增冯静汪建曹晓涵韩兴发
Owner SICHUAN AGRI UNIV