Molecular breeding method for thickening intermuscular bones of grass carps and black carps

A technology of molecular breeding and intermuscular spines, which is applied in the field of molecular breeding of thickened intermuscular spines of grass carp and herring, can solve problems such as affecting food processing and consumption, and achieve the effects of increasing ossification area and fast growth rate.

Pending Publication Date: 2020-08-18
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are a large number of intermuscular spines in the muscles of grass carp and herring, which affect food processing and people's consumption to a certain extent.

Method used

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  • Molecular breeding method for thickening intermuscular bones of grass carps and black carps
  • Molecular breeding method for thickening intermuscular bones of grass carps and black carps
  • Molecular breeding method for thickening intermuscular bones of grass carps and black carps

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Experimental program
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Effect test

Embodiment 1

[0060] (1) Cloning of partial nucleotide sequence of grass carp mstn gene

[0061] Download the mstn nucleotide sequences of several fishes that are closely related to grass carp in the gene database NCBI, and perform homology comparison, select relatively conservative regions to design PCR amplification primers, the primer sequences are:

[0062] Grass carp mstn:

[0063] F: 5'-GTGTATTAATTGCATGTGGTCC-3' (SEQ ID NO: 3);

[0064] R: 5'-GCTGTTACAGCAAAGATATAAATG-3' (SEQ ID NO: 4);

[0065] Using grass carp cDNA as a template, the mstn sequence was amplified to obtain a partial sequence of the coding region of grass carp mstn, as shown in SEQ ID NO:1.

[0066] (2) Construction of deletion mutant strains of grass carp mstn gene

[0067] The gene knockout method was used to obtain the grass carp mstn gene deletion mutant strain. Taking the method of constructing the deletion strain of crispr / cas9 as an example, the specific steps are as follows:

[0068] (i) Grass carp mstn gene knockout target ...

Embodiment 2

[0092] (1) Cloning of the nucleotide sequence of the mstn gene of herring

[0093] Download the herring mstn sequence from the gene database, design primers, and amplify the partial sequence of the coding region of mstn. The primer sequence is:

[0094] Herring mstn

[0095] F: 5'-TGCATGTGGTCCAGTGGGTTATG-3' (SEQ ID NO: 5);

[0096] R: 5'-CTCTTTGCCGTTGAAGTAAAG-3' (SEQ ID NO: 6);

[0097] Using herring cDNA as a template, the mstn sequence was amplified to obtain a partial sequence of the coding region of herring mstn, as shown in SEQ ID NO: 2.

[0098] (2) Construction of a mutant strain of herring mstn gene deletion

[0099] The gene knockout method was used to obtain a mutant strain of herring mstn gene deletion. Taking the method of constructing a deletion strain of crispr / cas9 as an example, the specific steps are as follows:

[0100] (i) Selection of herring mstn target genes

[0101] The herring mstn sequence has three exons: exon1, exon2, and exon3. In order to more efficiently perfo...

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Abstract

The invention discloses a molecular breeding method for thickening intermuscular bones of grass carps and black carps, and belongs to the technical field of aquatic organism breeding. The molecular breeding method provided by the invention is based on a technical means of gene editing, obtains an F0 generation of grass carp and black carp mstn genes subjected to targeted mutation, and obtains mutant individuals with mstn deletion and increased intermuscular bony area through passage. In the invention, a parent method for obtaining grass carp and black carp with increased intermuscular boning area by utilizing a gene editing technology is provided for the first time. According to the method, wild grass carps and black carps which have thick inherited intermuscular bones can be cultivated ona large scale in production, different from a transgenic method, the method can be applied to artificial breeding, overcomes the difficulty that intermuscular thorns are small and difficult to process in production, does not need to worry about the influence of transgenic food on people, is convenient for people to find the intermuscular thorns more easily when eating grass carps and black carps,and is easy to promote in production.

Description

Technical field [0001] The invention belongs to the technical field of aquatic product breeding, and specifically relates to a molecular breeding method for thickening the intermuscular spines of grass carp and herring. Background technique [0002] Grass carp and herring belong to the four major home fishes. They have the characteristics of short growth cycle and high survival rate during the breeding process. They are the main freshwater farmed fishes in my country. Their tender meat and delicious meat occupy a dominant position in freshwater fish farming in China. , Is a popular aquatic product in the market, which is deeply loved by consumers. The use of gene editing is an important technical means for the breeding of fish traits. Through gene editing technology, specific genes can be targeted to edit specific genes to obtain new varieties with excellent traits, which is of great significance to the aquaculture industry to breed varieties with excellent breeding performance. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/90C12N15/89C12N9/22A01K67/027A01K61/10
CPCC12N15/113C12N15/902C12N15/89C12N9/22C07K14/461C07K14/475A01K67/0276A01K61/10C12N2310/20A01K2207/15A01K2227/40A01K2267/02A01K2217/15A01K2217/075Y02A40/81
Inventor 鲍宝龙车金远
Owner SHANGHAI OCEAN UNIV
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