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Partial-base-deletion myostatin gene capable of being expressed in cattle body and application thereof

A myostatin and gene technology, applied in the field of genetic engineering, can solve problems such as hindering secondary structure and hindering gene sequence optimization

Inactive Publication Date: 2018-09-21
INNER MONGOLIA UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few naturally mutated MSTNs in animals, so how to base-knock out the original MSTN gene in mice so that it can play a role in mammals is an urgent technical problem in this field. There are many challenges in the process: firstly, how to obtain the gene sequence that can be expressed, and secondly, to construct the targeting vector. Considering the targeting efficiency of the targeting vector, the stability of the foreign gene mRNA, and the influence of the secondary structure of the mRNA on the translation efficiency, While optimizing, be sure to avoid the formation of secondary structures that hinder the expression of specific mRNAs
The above challenges hinder the optimization of gene sequences

Method used

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  • Partial-base-deletion myostatin gene capable of being expressed in cattle body and application thereof
  • Partial-base-deletion myostatin gene capable of being expressed in cattle body and application thereof
  • Partial-base-deletion myostatin gene capable of being expressed in cattle body and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] 1. MSTN target site design and synthesis

[0070] 1. Log in to the NCBI website, search for the bovine MSTN gene sequence published on the website and download it (Gene ID used in this experiment: JQ711180.1).

[0071] 2. Open the target prediction website (http: / / www.genome-engineering.org / crispr / ), open the online target design tool "CRISPR design tool" in the website, enter the gene sequence in the text box, and select the species to submit Then, all target sites of the sequence can be given, and potential off-target sites and comprehensive scores of each site can be given.

[0072] 3. According to the target site sequence given on the website, after comprehensive consideration of various parameters, two sets of target sites with lower predicted off-target effects are selected. Sites such as figure 1 Shown. Among them, A: sgRNA1, B: sgRNA2, C: Donor sequence. sgRNA is marked with the darkest color in the original sequence, the lightest color is marked with bases represen...

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Abstract

The invention discloses a partial-base-deletion myostatin gene capable of being expressed in a cattle body, in particular to an MSTN mutant gene. The MSTN mutant gene is formed by knocking out severalbase sequences of a first intron of an MSTN gene. The MSTN mutant gene can promote the occurrence of a double-muscle phenomenon of a cattle skeletal muscle. The invention further discloses application of the MSTN mutant gene.

Description

Technical field [0001] The present invention relates to genetic engineering technology in the field of biotechnology, in particular to a myostatin gene codon and its application, and more specifically to a myostatin gene with partial base deletion (ie MSTN mutation) that can be expressed in cattle. Gene) and its applications. Background technique [0002] MSTN is a myostatin gene that belongs to the TGF-β superfamily. Its synthesis is mainly completed by skeletal muscle. It belongs to the secreted type of polypeptides. As a member of the TGF-β superfamily, it has the same organisms as this family. Structure: including the secretion signal peptide at the N-terminus, proteolytic processing site (RSRR) and the mature peptide region at the C-terminus, containing a cystine knot structure, these components form a 52kDa The precursor protein without activity can form a 26kDa active peptide after processing through the intermediate proteolytic site, thus exerting its biological function...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/85A01K67/027
CPCC07K14/70575A01K67/0276A01K2217/075A01K2227/101A01K2267/02
Inventor 李光鹏魏著英白春玲高洋陈晨王东佟彬张立
Owner INNER MONGOLIA UNIVERSITY
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