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A method for promoting the proliferation of bovine skeletal muscle satellite cells in vitro

A satellite cell, in vitro proliferation technology, applied in the direction of using a vector to introduce foreign genetic material, cells modified by introducing foreign genetic material, recombinant DNA technology, etc. It can solve the problems of complex process and achieve the effect of strong in vitro proliferation ability.

Active Publication Date: 2015-10-14
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

[0003] The process of skeletal muscle proliferation and differentiation involves the expression of multiple genes, signaling pathways and network regulation, and the process is extremely complex

Method used

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  • A method for promoting the proliferation of bovine skeletal muscle satellite cells in vitro
  • A method for promoting the proliferation of bovine skeletal muscle satellite cells in vitro
  • A method for promoting the proliferation of bovine skeletal muscle satellite cells in vitro

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Embodiment Construction

[0025] 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.

[0026] Design idea of ​​the present invention is:

[0027] A pair of PCR primers were designed according to the bovine miR-133b gene sequence, XhoI restriction site and BamHI restriction site were introduced at the 5' ends of the two primers, and miR-133b containing miR-133b was amplified from the bovine genome using the pair of primers The full length of the gene is a sequence of 311bp, and the sequence is inserted into the vector pEGFP-C1 by using the introduced restriction site to construct the expression vector pEGFP-C1-miR133b of the bovine miR-133b gene; then use the obtained expression vector pEGFP- C1-miR133b transfected bovine skeletal muscle satellite cells, so that bovine skeletal mus...

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Abstract

The invention relates to a method for facilitating in vitro multiplication of a bovine skeletal muscle satellite cell. The method comprises the following steps: obtaining a target sequence containing a bovine miR-133b gene sequence; building an expression vector pEGFP-C1-miR133b; transfecting the bovine skeletal muscle satellite cell by the pEGFP-C1-miR133b. By application of the method for facilitating in vitro multiplication of the bovine skeletal muscle satellite cell, the in vitro multiplication ability of the bovine skeletal muscle satellite cell is improved by obtaining over-expression miniature RNA miR-133b of the bovine skeletal muscle satellite cell and using the effect of the miR-133b, and differentiation is effectively inhibited. The over-expression vector of the miR-133b is built by using the pEGFP-C1, and amplification and differentiation of a bovine muscle satellite cell are regulated by the miniature RNA, so as to obtain a high-purity bovine muscle satellite cell. A good cell model is provided for researching the function and the effect of each regulatory factor in amplification and differentiation processes of the muscle satellite cell.

Description

technical field [0001] The invention belongs to the cell engineering technology in the field of biotechnology, and in particular relates to a method for promoting the proliferation of bovine skeletal muscle satellite cells in vitro. Background technique [0002] Skeletal muscle satellite cells (skeletal muscle satellite cells) are myogenic stem cells with differentiation and proliferation potential in skeletal muscle. They usually exist between the muscle fiber sarcolemma and the basement membrane in a resting state, and can be activated under certain conditions. Proliferate and differentiate to form skeletal muscle cells. Muscle satellite cells play a very important role in the growth, development and regeneration of muscles in animals after birth. The activation, proliferation and differentiation of muscle satellite cells are regulated by many factors. Understanding the regulation mode of skeletal muscle satellite cells can increase the possibility of artificially controll...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N5/10
Inventor 丁向彬王轶敏刘新峰郭宏代阳聂永伟陈微微
Owner TIANJIN AGRICULTURE COLLEGE
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