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.
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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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