Method for introducing frame-shift mutation in MSTN (myostatin) genes of cattle

A frame-shift mutation and gene technology, applied in the field of genetic engineering, can solve problems such as inability to perform biological functions, and achieve the effects of abnormal muscle groups, tender meat, and thin skin

Inactive Publication Date: 2014-03-19
TIANJIN AGRICULTURE COLLEGE +1
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Problems solved by technology

This deletion is located at the 7th amino acid of the C-terminal region, which makes MSTN lose 102 amino acids. Therefore, a truncated protein is translated, which cannot exert its biological function of inhibiting muscle growth, thus showing double-muscle symptoms

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  • Method for introducing frame-shift mutation in MSTN (myostatin) genes of cattle
  • Method for introducing frame-shift mutation in MSTN (myostatin) genes of cattle

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Embodiment Construction

[0031] The present invention will be further described below in conjunction with specific embodiments, and its specific embodiments should be understood as illustrations only, not limiting, and the protection scope of the present invention cannot be limited by the following examples.

[0032] Design idea of ​​the present invention is:

[0033] According to the principle of applying the Cre-LoxP system to introduce frameshift mutations into the genome, two homology arms were designed, and the exon carrying the frameshift mutation was located on the short homology arm near the end of the screening gene. In order to keep the genome as unaffected as possible before and after targeting, the insertion point is selected in the intron, so that the inserted LoxP and other redundant sequences can be located in the intron. The frameshift mutation should be placed as close to the positive screening marker gene as possible. The frameshift mutation is located at the third exon of the bovine...

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Abstract

The invention discloses a method for introducing frame-shift mutation in bovine MSTN (myostatin) genes. According to the method, a targeting vector for the frame-shift mutation of the bovine MSTN genes is established, and the frame-shift mutation is introduced into the third exon of the bovine MSTN genes by virtue of homologous recombination. The method comprises the following steps of: synthesizing a homologous short arm containing the frame-shift mutation of MSTN, and allowing the homologous short arm to be short of 11bp on the third exon to form the frame-shift mutation; synthesizing a homologous long arm of the MSTN, and inserting a vector pFPC-1 by virtue of a restriction enzyme cutting site to establish the targeting vector pFPC-MSTN of the frame-shift mutation in the bovine MSTN genes; and transfecting bovine somatic cells after the linearization of the targeting vector, and introducing the frame-shift mutation in the bovine MSTN genes by virtue of the homologous recombination. Transgenic cattle with the frame-shift mutation of the MSTN genes can be obtained in the production by virtue of the cells, thereby improving beef production traits of the cattle and improving the quality and yield of beef.

Description

technical field [0001] The invention belongs to the genetic engineering technology in the field of biotechnology, and relates to a method for introducing a frameshift mutation into bovine MSTN gene through gene targeting technology. The frameshift mutation is caused by the deletion of a total of 11 bp from 4843bp to 4853bp of the MSTN gene. Background technique [0002] Growth / difference factor-8 (growth / difference factor-8, GDF-8) was discovered in 1997 by Mepherron and Lee of the John Hopkins University School of Medicine in the United States when studying the transforming growth factor beta (TGF-beta) superfamily of mice. A new protein factor gene, they named this gene growth / differentiation factor-8, namely GDF-8. Experimental studies have shown that GDF-8 normally encodes a protein that exhibits the activity of a negative regulator of skeletal muscle growth, so it is called myostatin (MSTN). [0003] The bovine MSTN gene is located between the centromere of chromosom...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/10
Inventor 郭宏李光鹏丁向彬刘新峰葛秀国李吉霞
Owner TIANJIN AGRICULTURE COLLEGE
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