Molecular breeding method for thickening intermuscular bones of silver carp and bighead carp

A molecular breeding and intermuscular thorn technology, applied in chemical instruments and methods, biochemical equipment and methods, and other methods of inserting foreign genetic materials, etc., can solve problems affecting food processing and consumption, and achieve increased ossification area and growth. fast effect

Pending Publication Date: 2020-08-07
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, silver carp and bighead carp have a large number of intermu

Method used

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  • Molecular breeding method for thickening intermuscular bones of silver carp and bighead carp
  • Molecular breeding method for thickening intermuscular bones of silver carp and bighead carp
  • Molecular breeding method for thickening intermuscular bones of silver carp and bighead carp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] (1) Cloning of partial nucleotide sequence of silver carp mstn gene

[0071] Download the mstn nucleotide sequences of several fish species that are closely related to silver carp in the gene database NCBI, and perform homology comparisons, select relatively conserved regions to design PCR amplification primers, and the primer sequences are:

[0072] Chub mstn:

[0073] F: 5'-GATGGGTAAAGCAGCCTTC-3' (SEQ ID NO: 3)

[0074] R: 5'-AGCAGAGAATGTCGTCCATTTC-3' (SEQ ID NO: 4)

[0075] Using silver carp cDNA as a template, the mstn sequence was amplified to obtain a partial sequence of the coding region of silver carp mstn, as shown in SEQ ID NO:1.

[0076] (2) Construction of deletion mutant strain of silver carp mstn gene

[0077] The mstn gene deletion mutant line of silver carp was obtained by gene knockout method, taking the method of constructing deletion line with CRISPR / Cas9 as an example, the specific steps are as follows:

[0078] (i) Silver carp mstn gene knockout...

Embodiment 2

[0103] (1) Cloning of the nucleotide sequence of the mstn gene of bighead carp

[0104] Download the mstn nucleotide sequence of fish with a close relationship with bighead carp from the gene database NCBI, and perform homologous comparison, select a relatively conserved region to design PCR amplification primers, and use wild bighead carp cDNA as a template to carry out mstn sequence The partial sequence of the coding region of bighead carp mstn was obtained by amplification, as shown in SEQ ID NO:2.

[0105] The amplification primers are:

[0106] F: 5'-GTATTAATTGCATGTGGTCCAG-3' (SEQ ID NO: 5)

[0107] R: 5'-CTCTTTGCCGTTGAAGTAAAG-3' (SEQ ID NO: 6)

[0108] (2) Construction of the deletion mutant strain of the bighead carp mstn gene

[0109] The gene knockout method was used to obtain the mstn gene deletion mutant strain of bighead carp. Taking the method of constructing the deletion strain with CRISPR / Cas9 as an example, the specific steps are as follows:

[0110] (i) Se...

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Abstract

The invention belongs to the technical field of aquatic organism breeding, and discloses a molecular breeding method for thickening intermuscular bones of silver carp and bighead carp. The molecular breeding method provided by the invention is based on a technical means of gene editing, an F0 generation of targeted mutation silver carp and bighead carp mstn (myostatin) genes is obtained, and mutant individuals with deleted mstn and increased intermuscular ossification areas are obtained through passage. In the invention, a parent method for acquiring silver carp and bighead carp with increasedintermuscular ossification areas by utilizing a gene editing technology is put forward for the first time. According to the method, wild silver carp and bighead carp which have thick intermuscular bones and can be inherited can be cultivated on a large scale in production; different from a transgenic method, the method can be applied to artificial breeding, the difficulty that intermuscular bonesare fine and small and difficult to process in production is overcome, people do not need to worry about the influence of transgenic food on people, people can find intermuscular bones more easily when eating silver carp and bighead carp, and the method is easy to popularize in production.

Description

technical field [0001] The invention belongs to the technical field of aquatic organism breeding, and in particular relates to a molecular breeding method for thickening intermuscular spines of silver carp and bighead carp. Background technique [0002] Silver carp and bighead carp belong to the four major family fishes. They have the characteristics of short growth cycle and high survival rate in the breeding process. They are important freshwater cultured fish in my country. 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 for the aquaculture industry to cultivate varieties with excellent breeding performance. At present, many domestic researchers have carried out research on molecular breeding such as body color, disease resistance, and gonad development of farmed fish t...

Claims

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Application Information

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