Activation method for producing transgenic cloned pigs

Abstract

The invention discloses an activation method for producing transgenic cloned pigs. The combined activation of electrical activation and chemical activation is adopted, and actidione is firstly used for activation treatment for 3min-25min before the adoption of the electrical activation. Preferably, the activation treatment time by the actidione is 10-20min; further preferably, the activation treatment time by the actidione is 10min; and the concentration of the actidione for activation treatment is 5-10 micrograms / mL. According to the activation method disclosed by the invention, by changing the sequence of a common combined activation way, a scheme of firstly performing chemical activation and then performing electrical activation is adopted, the chemical-assisted activation time in the activation scheme is shortened, the embryonic development capability which is similar to an original combined activation scheme of firstly performing electrical activation and then performing chemical activation, with long activation treatment time can be obtained, and low blastocyst apoptosis rate and DNA (deoxyribonucleic acid) methylation level during a blastocyst stage can be further obtained. The transgenic cloned pigs can be successfully produced after transplantation of an embryo produced by adopting the scheme.

Description

[0001] technical field [0002] The invention relates to the production of transgenic cloned pigs, in particular to an activation method for producing transgenic cloned pigs. Background technique [0003] One of the main reasons for the inefficiency of cloning and abnormal development is the abnormality in the apparent level of cloned embryos, such as the methylation level of the whole genome. [0004] The activation of cloned embryos is the key to start the development of cloned embryos. Common activation methods for cloned embryos include physical activation (such as electrical pulse stimulation, mechanical stimulation, temperature change, etc.) and chemical activation (such as calcium ionophore, ethanol, protein synthesis inhibitors, protein kinase inhibitors, etc.). Although the methods are different, the activation mechanism is basically the same, which is to restore the division of oocytes that have stopped at the MII stage. Chemical activation has a poor ability to ...

AI Technical Summary

Problems solved by technology

[0005] In the existing activation scheme, the activation scheme of using CHX for 4-6 hours after electrical activation is usually adopted. In this scheme, the transgenic cloned embryos should be treated with the chemical auxiliary activator CHX for 4-6 hours. CHX exposure for a long time will cause an increase in the apoptosis rate of the embryo and an excessively high level of global methylation, which may affect the cloning efficiency and the health of the offspring, and it will take a lot of time and labor costs

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