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Kit for preparing transgenic animals through testis injection

A transgenic animal and testicular injection technology, applied in the field of kits, can solve the problems of inability to obtain living offspring and low efficiency of transgenic animals, and achieve the effects of improving transgenic efficiency, low toxicity, and animal safety

Inactive Publication Date: 2013-01-16
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficiency of transgenic animals obtained by testicular injection is low, and living offspring are often not obtained

Method used

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  • Kit for preparing transgenic animals through testis injection
  • Kit for preparing transgenic animals through testis injection
  • Kit for preparing transgenic animals through testis injection

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Experimental program
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Effect test

preparation example Construction

[0030] Preparation method of magnetosome (BMP): it can be prepared by existing method. Specifically, magnetosomes (BMPs) can be prepared according to the published national patent (publication number: CN101376900; patent name: a method for high-density culture of magnetotactic bacteria with high yield of magnetosomes).

[0031] The plasmid DNA with the reporter gene green fluorescent protein (GFP) was purchased from Shanghai Gemma Pharmaceutical Technology Co., Ltd., the product catalog number is F-07.

Embodiment 1

[0032] Embodiment 1, preparation transgenic animal

[0033] 1. Preparation of BMPS-PEI-DNA complex:

[0034] Take an appropriate amount of BMP and mix briefly for 10-20s in an ultrasonic vibrator, and then quickly place it on ice. -20s. Then add the plasmid DNA (BMPS:PEI:plasmid DNA=0.5:0.5:1, g / g / g) with the reporter gene green fluorescent protein (GFP) and mix thoroughly again, then use the glass injection needle to draw an appropriate amount of BMPs- The PEI-DNA mixture is ready for use.

[0035] 2. Injection of BMPs-PEI-DNA mixture in the testes of male animals:

[0036] The male animals were anesthetized and fixed, and the surgical department was disinfected. Open the abdominal cavity, find the testes and pull them out of the abdominal cavity to fix the testes. Puncture the testicular capsule with a 1 mL sterile syringe needle (22G) along the longitudinal axis of the testis. Then, the glass needle absorbed with the BMPs-PEI-plasmid DNA mixture was inserted into the ...

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Abstract

The invention discloses a kit for preparing transgenic animals through testis injection. The kit for preparing transgenic animals comprises a composition composed of polyethyleneimine and magnetosomes. The kit has the advantages of substantial improvement of the transgenic efficiency, and obtaining of a large amount of transgenic live progenies. The composition in the kit has the advantages of small toxicity, and safety to animals. So the use of the kit in transgenic operation improves the production efficiency of the transgenic animals, and is of great importance in the transgenic animal preparation field.

Description

technical field [0001] The present invention relates to a kit for preparing transgenic animals by testicular injection. Background technique [0002] The testicular injection method is to inject exogenous DNA directly into the testes of male animals, so that the target gene can be transfected into the germ cells at various stages of development, and the sperm transfected with the target gene can be matured in vivo to obtain transgenic animals by natural mating or artificial insemination . It is characterized by simple operation, low cost and suitable for mass production of transgenic animals. However, the efficiency of transgenic animals obtained by testicular injection is low, and living offspring are often not obtained. [0003] Polyethyleneimine (PEI) has a high cationic charge density and has a strong "proton sponge effect", which can effectively destroy lysosomes and quickly release the loaded nucleic acid from the endocytosis pathway, and can be used as Gene carrier...

Claims

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Application Information

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IPC IPC(8): C12N15/89A01K67/027
Inventor 王超李颍唐涛杨蕾蕾池亚菲余辰陈振文
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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