Method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5

A satellite cell and myogenic differentiation technology, applied in the cell and biological fields, can solve the problems of undiscovered patent publications, etc., and achieve the effect of promoting the myogenic differentiation ability and improving the myogenic differentiation ability.

Active Publication Date: 2020-04-07
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

[0004] Through the search, no patent publications related to the patent application of the present invention have been found

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  • Method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5
  • Method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5
  • Method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5

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Embodiment Construction

[0024] The embodiments of the present invention will be described in detail below. It should be noted that the embodiments are illustrative, not restrictive, and cannot limit the protection scope of the present invention.

[0025] The raw materials used in the present invention, unless otherwise specified, are conventional commercially available products; the methods used in the present invention, unless otherwise specified, are conventional methods in the art.

[0026] A method for promoting bovine skeletal muscle satellite cell myogenic differentiation by overexpressing PSMB5, ​​the steps are as follows:

[0027] (1) Acquisition of the target sequence containing the bovine PSMB5 gene sequence:

[0028] Design a pair of PCR primers according to the bovine PSMB5 gene sequence, introduce XhoI restriction site and XbaI restriction site at the 5' ends of the two primers, and use the pair of primers to amplify the CDS region containing the PSMB5 gene from the bovine genome Sequen...

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Abstract

The invention relates to a method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5. The method comprises the following steps: (1) obtaining atarget sequence containing a bovine PSMB5 gene sequence; (2) constructing an expression vector pcDNA3.1-PSMB5; (3) transfecting bovine skeletal muscle satellite cells by pcDNA 3.1-PSMB5, using the obtained expression vector pcDNA3.1-PSMB5 for transfecting bovine skeletal muscle satellite cells by adopting a liposome transfection reagent so that the bovine skeletal muscle satellite cells overexpress the PSMB5, and improving the in-vitro differentiation capacity of the bovine skeletal muscle satellite cells by utilizing the effect of the PSMB5. According to the method, PSMB5 is used for regulating and controlling proliferation and myogenic differentiation of bovine muscle satellite cells, the enlightenment can be provided for research on the ubiquitination process of muscle development anddifferentiation, and a new thought and reference are provided for clinical research and diagnosis and treatment of muscle development and differentiation and injury repair.

Description

technical field [0001] The invention belongs to the field of biology and cell technology, in particular to a method for promoting bovine skeletal muscle satellite cell myogenic differentiation by overexpressing PSMB5. Background technique [0002] Skeletal muscle satellite cells are myogenic stem cells with the potential of proliferation and differentiation in skeletal muscle. They usually exist between the muscle fiber sarcolemma and 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 controlling the formation of muscle cells. [0003] T...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/57C12N5/10
CPCC12N15/85C12N9/6427C12N5/0659C12Y304/21001C12N2800/107C12N2510/00
Inventor 王轶敏来裕婷丁向彬张林林李新郭益文郭宏
Owner TIANJIN AGRICULTURE COLLEGE
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