Treatment method for preparing sperm of transgenic mouse

Abstract

The invention belongs to a transgenic field, and relates to a treatment method for preparing sperm of a transgenic mouse. The treatment method for preparing sperm of the transgenic mouse, comprises: incubating mouse epididymal sperm at 37 DEG C in a 5% CO2 incubator for 8-10min, mixing a linear DNA fragment containing a target gene into the sperm, ice-bathing the sperm at 2-5 DEG C for 25-30min, heat-shocking the sperm at 40-45 DEG C for 1-3min, ice-bathing the sperm at 2-5 DEG C for 3-5min, and then incubating the sperm at 37 DEG C in a 5% CO2 incubator for 50-70min for capacitation The method can raise permeability of a sperm membrane, is convenient for introducing a gene fragment into the sperm, and raises transgenic efficiency. The invention solves the problems of low efficiency, bad stability and difficult passage when sperm is used to prepare the transgenic mouse. An effective method for preparing the transgenic mouse is disclosed.

Description

technical field

[0001] The invention belongs to the field of transgenics, and relates to a sperm treatment method for preparing transgenic mice. Background technique

[0002] Transgenic methods have been widely used in various fields of modern life science research and gene function research. At present, gene transfer techniques mainly include DNA microinjection, ES cell-mediated method, retroviral vector method, sperm carrier method and so on. Although these methods can be used to introduce foreign genes into target cells and obtain corresponding transgenic animals, there are some limitations. Among them, the most commonly used DNA microinjection method requires expensive experimental equipment, cumbersome work, high technical requirements, low rate of exogenous gene introduction, and high production costs; although the gene transfer efficiency of the retroviral vector method is high, the gene insertion It is random, and the size of the DNA fragments carried is limited, a...

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