Mechano growth factor polypeptide, preparation method and application thereof

A technology of growth factors and engineering bacteria, applied in growth factors/inducing factors, biochemical equipment and methods, botany equipment and methods, etc., can solve problems such as difficult to obtain pure products, unstable MGF, unsatisfactory results, etc. Achieve the effects of promoting cell proliferation, simple preparation method, and good biological activity

Inactive Publication Date: 2010-11-24
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to obtain MGF, experts at home and abroad have carried out some research work, but the results are not ideal. Because MGF is unstable, it is easy to form inclusion bodies when expressed in engineering bacteria, and it is difficult to obtain pure products.

Method used

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  • Mechano growth factor polypeptide, preparation method and application thereof
  • Mechano growth factor polypeptide, preparation method and application thereof
  • Mechano growth factor polypeptide, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0018] MGF 41 Construction of expression vector

[0019] Using the pExSecI-MGF stored in the laboratory as a template, use specific primers: Forward primer: gat at gga tcc gctagc gct cgc tct gtc cgt gcc cag and Reverse primer: cgc gc ctc gag tta ctt gtg ttc ttc aaa tgt ac for PCR amplification The resulting amplified product was recovered by the OMEGA gel extraction kit to recover the target gene, and then digested with Nhe I and Xho I, and the digested product was ligated with the vector pRSETc that had undergone the same double digestion with T4 DNA ligase. The ligation product was transformed into Escherichia coli DH5α, and positive clones were picked for colony PCR and enzyme digestion identification. DNA sequencing analysis showed that MGF 41 It has been cloned into pRSETc, and the recombinant plasmid is named pRSETc-MGF 41 .

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Abstract

The invention provides a mechano growth factor polypeptide, a preparation method and an application thereof. The mechano growth factor polypeptide is the polypeptide of 41 amino acids at C end of a mechano growth factor, and the polypeptide is taken as an active region of the mechano growth factor. The preparation method comprises the following steps: cloning a nucleotide sequence of the polypeptide into an expression vector pRSETc to obtain a recombinant expression plasmid pRSETc-MGF41; transfecting escherichia coli with the plasmid to obtain an engineering strain for expressing the mechano growth factor polypeptide; and finally culturing and purifying the engineering strain to obtain the mechano growth factor polypeptide. The polypeptide has the function of promoting cell proliferation, thus the polypeptide can be used for preparing a medicine for promoting cell proliferation.

Description

technical field [0001] The present invention relates to growth factor, specifically belongs to force growth factor polypeptide (MGF 41 ) and its preparation method and use. Background technique [0002] Mechano growth factor (MGF) is a regulator of local muscle growth caused by mechanical stretch discovered by Goldspink et al. Under the stimulation of muscle force, the insulin-like growth factor I (Insulin-like growth factor, IGF-I) gene produces two isomers through alternative splicing, one is similar to the liver type IGF-I, named muscle Type IGF-I (IGF-I Ea); the other is named force growth factor (MGF) because it is very sensitive to mechanical stimulation. Studies have found that MGF is not only a growth factor, but also a repair factor. The function of MGF is mainly related to its C-terminal domain. MGF is uncommon under normal physiological conditions, but when skeletal muscle or cardiac muscle is under increased mechanical load, its expression increases, and it p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475C12N15/12C12N15/63C12N15/70C12N1/21A61K38/18A61P21/00
Inventor 宋莉董俊丽李卓玉祝文思刘晓辉
Owner SHANXI UNIV
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