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Production method of recombination fibroblast growth factor-8b and application thereof

A technology of fibroblasts and growth factors, applied in the field of genetic engineering, to achieve the effect of fast and convenient acquisition and promotion of cell proliferation

Inactive Publication Date: 2016-03-02
WENZHOU MEDICAL UNIV +1
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  • Abstract
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Problems solved by technology

[0005] One of the purposes of the present invention is to provide a production method for recombinant fibroblast growth factor-8b protein, to solve the problem of being able to produce biologically active FGF8b protein in large quantities, to meet the needs of production and FGF8b protein

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  • Production method of recombination fibroblast growth factor-8b and application thereof
  • Production method of recombination fibroblast growth factor-8b and application thereof
  • Production method of recombination fibroblast growth factor-8b and application thereof

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Embodiment Construction

[0046] In accordance with the present invention, conventional molecular biology, microbiology, cytology and DNA recombinant techniques within the skill of the art may be employed. Definitions of terms where they appear in this document are as follows.

[0047] "DNA molecule" refers to the polymeric form of deoxyribonucleotides (thymine, cytosine, adenine or guanine), which are the main constituents of chromosomes and the material that makes up genes. This term refers only to the primary and secondary structure of the molecule, without restricting it to any specific tertiary form. The term includes double-stranded DNA found especially in linear DNA molecules, viruses, chromosomes, plasmids. The structure discussed here is conventionally given only the sequence of the sense strand of DNA in the 5' to 3' direction.

[0048] A "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been ligated. One type of vector is a "plasmid," w...

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Abstract

The invention discloses a production method of a recombination fibroblast growth factor-8b protein. The method comprises the following steps that 1, an insect baculovirus expression vector system is utilized for expressing a fibroblast growth factor-8b protein in an insect cell, wherein the recombination baculovirus Bacmid-FGF8b is obtained, genes of the FGF8b are fibroblast growth factor-8b genes, and the base sequence of the FGF8b is shown in the SEQ ID No:1; the Bacmid-FGF8b recombination baculovirus of the certain MOI obtained in the first step is used for infecting insect cells, and vibration and suspension are performed at the temperature of 25-29 DEG C for culture for 36-120 h after infection is performed to obtain a cell sap; 2, the fibroblast growth factor-8b protein is obtained by purifying the cell sap obtained in the first step. The invention further discloses application of the fibroblast growth factor-8b protein for treating the Parkinson disease.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a production method and application of recombinant fibroblast growth factor-8b. Background technique [0002] Fibroblast growth factor-8 (fibroblast growth factor 8, FGF-8) is a member of the fibroblast growth factor family, and has 30% to 40% homology with known family members. FGF8 was first extracted from the androgen-dependent mouse breast cancer cell line SC-3. The human FGF8 gene is located in the chromosome 10q24 segment, has 6 exons, and is composed of 233 amino acids. The mature FGF8 is a polypeptide composed of 194 amino acid residues (molecular weight is about 23kDa) by removing the N-terminal 39 amino acid signal peptide. It is a basic protein with an isoelectric point of about 10.5. Human FGF8 has four subtypes, a, b, e, and f, and FGF8b is its dominant expression type, and its binding ability to FGF receptors is the strongest. Extensive literature reports that ...

Claims

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Application Information

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IPC IPC(8): C12N15/866C07K14/50A61P25/16
Inventor 姜潮马吉胜田海山刘敏陈娜子
Owner WENZHOU MEDICAL UNIV
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