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Recombinant soluble human FGFR2 extracellular fragment and production method and application thereof

A technology of FGFR2 and production methods, applied in the direction of biochemical equipment and methods, applications, botany equipment and methods, etc., can solve problems such as carcinogenesis, cell transformation, and craniosynostosis, and achieve reduced levels, good activity, and production The effect of simple process

Inactive Publication Date: 2008-03-12
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, the gene mutation of FGFR can lead to changes in the affinity of FGFR to FGFs, thereby causing bone developmental deformities, such as achondroplasia, acral developmental deformities such as syndactyly, craniosynostosis, etc.; Overexpression of FGFR or misregulation of alternative splicing of mRNA can lead to cellular transformation and carcinogenesis

Method used

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  • Recombinant soluble human FGFR2 extracellular fragment and production method and application thereof
  • Recombinant soluble human FGFR2 extracellular fragment and production method and application thereof
  • Recombinant soluble human FGFR2 extracellular fragment and production method and application thereof

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Embodiment 1

[0040] Construction of double mutants in the extracellular segment of FGFR2:

[0041] 1. Construction of the wild-type extracellular segment of FGFR2:

[0042]According to the amino acid sequence of the wild-type extracellular segment of the truncated FGF receptor, four pairs of primers were designed according to the codon preference principle of Escherichia coli, respectively marked as: upstream primers F1, F2, F3, F4; R4, as shown below, and Nde I and BamH I restriction endonuclease sites were introduced at the 5' ends of primers F4 and R4, respectively, to facilitate directional cloning of the gene. Each strip ranges from 105 to 106bp, and the full-length synthesis is 707bp. The reading frame contains a 684bp truncated version of the extracellular segment of FGFR2IIIC, starting from 150aa to 365aa, and adjacent to each strip. There is a 20bp overlap between the primers. There are four rounds of synthesis. In the first round, the innermost primers F1 and R1 are used for ov...

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Abstract

The present invention discloses a recombinant soluble FGFR2 extracellular domain and the production method as well s the application of the extracellular domain. The present invention is based on the high-efficient expression FGFR2 of escherichia coli. Qualitative and quantitative researches have done concerning a plurality of factors that affect the renaturing efficiency. A proper method for recombinant soluble FGFR2 extracellular domain is determined. The renaturing efficiency can be of 10 percent to 20 percent. The present invention is of low cost and short cycle. The present invention is suitable for mass production. And the present invention takes advantage of the combination of dissociative extracellular domain of FGFRs with the FGFs in the tumor tissue to reduce the amount of dissociative FGFs, competitively leading to the repressive FGFs signals and the restraining proliferation and transfer of tumor as well as the depressed formation of blood vessels. S252W+P253R extracellular domain and double-mutant FGFR2 extracellular domain are of the best efficiency.

Description

technical field [0001] The invention relates to the technical field of human FGF receptors, in particular to a recombinant soluble human FGFR2 extracellular segment and its production method and application. Background technique [0002] Fibroblast growth factors (Fibroblast growth factors FGFs) are a group of homologous heparin-binding polypeptides, and its family contains 23 members. FGFs have a wide range of biological activities, such as regulating cell proliferation, migration and differentiation in embryonic development, inducing wound repair, tumor growth and differentiation, and inducing neovascularization in adults. FGFs are potent mitogens for a variety of cell types. The diverse biological activities of FGFs are realized through the high-affinity receptors FGFRs on the cell surface. [0003] Both FGF and FGFR are important proto-oncogenes, which play a very important role in the physiological activities of tumor proliferation, angiogenesis, metastasis and apopto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12C12N1/21A61K36/17A61P35/00
Inventor 洪岸汪炬张志成
Owner JINAN UNIVERSITY
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