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Tri-cistron muscle specific two-way co-expression gene transferant and preparation method

A co-expression gene and tricistronic technology, applied in the field of genetic engineering, can solve problems such as mutual influence

Inactive Publication Date: 2015-11-18
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Unidirectional co-expression of multiple genes, often interacting

Method used

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  • Tri-cistron muscle specific two-way co-expression gene transferant and preparation method
  • Tri-cistron muscle specific two-way co-expression gene transferant and preparation method

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Embodiment Construction

[0086] In order to make the objects and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0087] The embodiment of the present invention provides a gene sequence of a tricistronic muscle-specific bidirectional co-expression gene transfer body: SEQ ID NO: 13, such as figure 1 As shown, the gene transfer body includes the MARs bovine nuclear matrix binding region from the 5' end to the 3' end, and the third exon (926-947 site) of bovine myostatin (MSTN) as the target Small hairpin RNA (shRNA), MRF4, MRF4 are optimized synthetic muscle-specific promoters, FAD3 is a humanized n-3 desaturase gene, IRES is an internal ribosome entry site, DsRed red fluorescent protein gene, SV40polyA is a terminator signal area. The independent gene tran...

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Abstract

The invention discloses a tri-cistron muscle specific two-way co-expression gene transferant and a preparation method thereof. The gene transferant comprises an MARs cow matrix attachment region, short hairpin RNA (shRNA) taking the third exon (926-947 loca) of cow muscle colyone as a target spot, an optimally synthesized muscle specific promoter (MRF4), a humanization n-3 desaturase gene (FAD3), an internal ribosome entry site (IRES), a red fluorescent protein gene (DsRed) and a termination signal region (SV40polyA), and the independent gene transferant does not have a pronucleus main vector sequence. According to the tri-cistron muscle specific two-way co-expression gene transferant and the preparation method, an RNA interference technology is firstly combined with the muscle specific overexpression FAD3 and DsRed, therefore, gene knock-down and inverse overexpression of related genes can be performed simultaneously, and a novel thought and approach can be supplied for safely and efficiently improving characters, such as muscle and disease related characters, controlled by polygenes.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering in the field of biotechnology, and relates to a tricistronic muscle-specific bidirectional co-expression gene transfer body and a preparation method thereof Background technique [0002] High-efficiency expression of foreign genes in transgenic animals must rely on good expression vectors. There is a greater need for multi-gene, tissue-specific co-expression vectors. [0003] MSTN, also known as myostatin, is a member of the transforming growth factor β superfamily. In 1997, McPherron and Lawler first prepared MSTN knockout mice [1] , confirming that its most important function is to negatively regulate the growth and development of skeletal muscle. As a secreted protein, MSTN is mainly produced by skeletal muscle. In different species, MSTN gene consists of 3 exons (exon1 / 2 / 3) and 2 introns (intron1 / 2). In mammals The mature peptide sequence homology of MSTN is very high, for examp...

Claims

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Application Information

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IPC IPC(8): C12N15/79C12N15/66
Inventor 李光鹏郭晶广璐张英白春玲魏著英扈廷茂
Owner INNER MONGOLIA UNIVERSITY
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