Recombinant HEK293 cell with high FGFR4 expression and application thereof

A high-expression technology of FGFR4, applied in the field of recombinant cells, can solve problems such as cell proliferation regulation disorder, and achieve the effect of improving specificity and sensitivity

Inactive Publication Date: 2013-07-10
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is known that about 50% of the expression products of oncogenes that lead to tumorigenesis and development have protein tyrosine kinase activity, and their abnormal expression will lead to the disorder of cell proliferation regulation, and then lead to tumorigenesis

Method used

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  • Recombinant HEK293 cell with high FGFR4 expression and application thereof
  • Recombinant HEK293 cell with high FGFR4 expression and application thereof
  • Recombinant HEK293 cell with high FGFR4 expression and application thereof

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Experimental program
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Embodiment Construction

[0028] On the basis of constructing the eukaryotic expression vector pcDNA3.1(+) / FGFR4 of FGFR4 full-length gene, the present invention transfects HEK293 cells to obtain a stable and high-expression FGFR4 recombinant HEK293 / FGFR4 cell line. The detailed description is intended to explain rather than limit the present invention.

[0029] The present invention uses pcDNA3.1(+) as the base carrier, and specifically selects HindIII and XhoI restriction sites as multiple cloning sites for connecting foreign genes.

[0030] 1) FGFR4 gene cloning

[0031] Select the pOTB7 / FGFR4 plasmid containing the full-length cDNA sequence of the FGFR4 gene as a template for cloning, use Primer Premier5.0 software to design corresponding specific primers, and add HindIII / XhoI restriction sites at both ends:

[0032] FGFR4-HindIII: 5'-AGT AAGCTT ATGCGGCTGCTGCTGGCCCT-3'

[0033] FGFR4-XhoI: 5'-GAT CTCGAG TCATGTCTGCACCCCAGACC-3'

[0034] LA Taq from Takara Company was used for amplification. T...

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Abstract

The invention discloses a recombinant HEK293 cell with high FGFR4 expression and application thereof. In the recombinant cell, an HEK293 cell serves as a host cell; and an exogenous expression vector for transfecting the host cell is an expression vector which comprises an FGFR4 full-length gene. The recombinant HEK293 cell with high FGFR4 expression is applied to screening of antitumor medicaments.

Description

technical field [0001] The invention belongs to the technical field of recombinant cells, and relates to a recombinant HEK293 cell highly expressing FGFR4 and an application thereof. Background technique [0002] Receptor tyrosine kinases (RTKs) are the largest class of enzyme-linked receptors, which are both receptors and enzymes, capable of binding to ligands and phosphorylating tyrosine residues of target proteins. All RTKs are composed of three parts: an extracellular domain containing a ligand-binding site, a single-transmembrane hydrophobic α-helical region, and an intracellular domain containing tyrosine protein kinase (RTK) activity. Receptor tyrosine kinase exists as a monomer and has no activity when it is not combined with a signal molecule; once a signal molecule binds to the extracellular domain of the receptor, two monomeric receptor molecules form a binary receptor on the membrane. In the aggregate, the tails of the intracellular domains of the two receptors ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12Q1/02
Inventor 贺浪冲张涛王嗣岑侯晓芳李淼杜晖
Owner XI AN JIAOTONG UNIV
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