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Transgenic method of marine flounder

A transgenic, seawater technology, applied in biochemical equipment and methods, cells modified by introducing foreign genetic material, veterinary instruments, etc., can solve the problems of lack of effective transgenic methods and inability to conduct large-scale transgenic research

Active Publication Date: 2013-10-09
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0003] The purpose of the present invention is to provide a transgenic method for marine flounder and flounder, that is, a method for gene transfer of flounder and flounder such as semi-smooth tongue sole without microinjection of fertilized eggs, thereby solving the lack of effective methods for flounder and flounder such as half-smooth tongue sole. Transgenic methods, the difficulty of large-scale transgenic research

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Embodiment Construction

[0023] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0024] 1. Construction of transgenic plasmids:

[0025] Using the VASA gene promoter specifically expressed in fish gonads or the CMV virus gene promoter expressed in the whole body and other gene promoters, the sex-related genes or other gene sequences of half-smooth tongue sole are inserted downstream of the promoters to construct transgenic plasmids.

[0026] The following takes the construction of the DMRT1 gene transgenic plasmid of half-smooth tongue sole driven by the red sea bream VASA gene promoter as an example to describe in detail.

[0027] (1) Gene elements: The plasmid pPmvasGFP with red sea bream vasa promoter and enhanced green fluorescent protein gene was used as the initial vector for the construction of the transgenic vector. The construction of the plasmid promoter was carried out according to the method of Lin et al. (2011...

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Abstract

The invention aims at providing a transgenic method of a marine flounder. The transgenic method of the marine flounder comprises the following steps of: transferring plasmids with exogenous genes into pubescent testis of a milter; and collecting seminal fluid of the transgenic milter and fertilizing with mature eggs of a hen, so as to transfer the exogenous gene into the mature eggs through sperms. The method avoids the problem that lots of embryos are dead due to micro-injection to the oosperm. By adopting the method, the exogenous gene can be effectively transferred into the oosperm of the flounder; the integration of the exogenous gene integrated in a transgenic fry is 12 to 21%; a novel method is provided for the study on gene transfer of the flounder; the problem that the transgenosis is hardly carried out on the oosperm of the flounder by micro-injection is solved; and novel technical means are provided for breeding in the flounder genetic engineering.

Description

technical field [0001] The invention belongs to the technical field of seawater fish genetics and breeding, and in particular relates to a transgenic method for seawater flounder and flounder, that is, a method for efficiently transferring exogenous genes to seawater flounder and flounder such as semi-smooth tongue sole. Background technique [0002] Gene transfer is an important means to develop high-yielding, disease-resistant new varieties of animals and plants, and it is also an effective method to analyze gene functions. Fish has been widely concerned because of its large number of fecundity, in vitro development of fertilized eggs, and easy operation. In 1985, Zhu Zuoyan from the Institute of Hydrobiology of the Chinese Academy of Sciences microinjected the exogenous growth hormone gene into the fertilized eggs of crucian carp and achieved success. Since then, countries around the world have done a lot of research on transgenic fish and made great progress. Microinje...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N5/10C12Q1/68A61D19/02
Inventor 陈松林胡乔木刘洋王娜
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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