Electroporation transfection method of eyespot larvae of mytilus coruscus

Abstract

The invention belongs to the field of aquatic product biogenetic engineering, and discloses an electroporation transfection method of eyespot larvae of mytilus coruscus. siRNA is designed according toa full-length sequence of thyroid hormone receptor genes of the mytilus coruscus obtained by clone, and different parameter settings are adopted to analyze the knock reduction situations of differentpulse patterns on the thyroid hormone receptor genes; the larvae after 96 hours of electroporation transfection are collected, and larvae RNA is extracted; and thyroid hormone receptor gene primers are designed, and the expression situation of the thyroid hormone receptor genes is detected by a fluorogenic quantitative PCR. According to the electroporation transfection method of the eyespot larvae of the mytilus coruscus, compared with the treatment without siRNA and non-sense siRNA, results show that the addition of the thyroid hormone receptor gene siRNA significantly reduces the expressionquantity of the genes, and the knock reduction efficiency is nearly 100%.

Description

technical field [0001] The invention belongs to the field of aquatic biological genetic engineering, and in particular relates to an electroporation transfection method for eye-spotted mussel larvae. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] Thick shelled mussel Mytilus coruscus is a seawater economic mollusk widely distributed in the coastal areas of my country, belonging to the mollusc phylum Bivalvia, and is an important aquaculture economic shellfish in China. Shengsi County, Zhejiang Province, my country is the main production area of ​​thick shelled mussels . Thick-shelled mussels are large and plump. They are bivalve seafood containing high-quality protein and have high nutritional value. They are deeply loved by people in coastal areas of our country. [0004] The supply of seedlings of mussels is mainly from artificial breeding. Larval metamorphosis is a very important stage in artific...

AI Technical Summary

Problems solved by technology

At present, the problems existing in the artificial propagation technology of thick-shelled mussels have not been solved, and the survival rate of seedlings and the quality of seedlings have been unstable, resulting in a shortage of seedlings.

There are no reports in shellfish, nor correspondingly established methods for relying on electroporation in shellfish larvae;

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