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FGFR1 high-expression recombinant HEK293 cell and application thereof

A recombinant cell, high-expression technology, which can be used in the determination/inspection of cells and microorganisms modified by the introduction of foreign genetic material, biochemical equipment and methods, etc. There are no reported problems in the application to achieve the effect of improving sensitivity and specificity

Inactive Publication Date: 2013-04-17
XI AN JIAOTONG UNIV
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Problems solved by technology

At present, there are many studies on the FGFR1 gene, but most of them only focus on the extracellular region of the receptor, or the construction of the carrier of some genes in the intracellular region, and there is no research on the construction of the full-length sequence FGFR1 carrier, the study of its biological characteristics and its application. to report

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  • FGFR1 high-expression recombinant HEK293 cell and application thereof
  • FGFR1 high-expression recombinant HEK293 cell and application thereof
  • FGFR1 high-expression recombinant HEK293 cell and application thereof

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Embodiment Construction

[0034] In the present invention, on the basis of constructing eukaryotic expression carrying pcDNA3.1(+)-FGFR1 of the full-length FGFR1 gene, HEK293 cells are transfected to obtain a recombinant HEK293-FGFR cell line with stable and high expression of FGFR molecules. The following is a detailed description of this aspect, which is an explanation of the present invention rather than a limitation.

[0035] The present invention is based on pcDNA3.1 (+) carrier (from the First Affiliated Hospital of Xi'an Jiaotong University donated), its plasmid map is as follows figure 1 As shown, the present invention selects the BamHI restriction site as the multiple cloning site for connecting foreign genes; the BamHI and EcoR I restriction sites are used as the detection site for vector construction.

[0036] 1) Construction of eukaryotic expression vector pcDNA3.1(+)-FGFR1

[0037] a. Design a pair of primers comprising restriction sites, and use the pCMV-SPORT6-FGFR1 plasmid (containing ...

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Abstract

The invention discloses an FGFR1 high-expression recombinant HEK293 cell and application thereof, wherein the recombinant cell takes an HEK293 cell as a host cell, and an exogenous expression vector for transfecting the host cell contains an FGFR1 full-length gene. Compared with the existing recombinant cell which only contains an FGFR1 extracellular region or is constructed by part of gene sequences, the surface of the recombinant HEK293-FGFR1 cell strain containing the FGFR1 full-length gene constructed in the invention can stably express FGFR1 molecules and has complete molecular activity.A cell membrane chromatography is adopted to detect the HEK293-FGFR1 cell, wherein gefitinib is remained to a certain degree, which can be applied to drug screening with FGFR as a target point.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the construction of non-cancer cells with high FGFR expression, in particular to a recombinant HEK293 cell with high FGFR1 expression based on an exogenous recombinant plasmid and its application. technical background [0002] Receptor tyrosine kinases (RTKs) are a huge family that play a very important role in the life activities of organisms. When the ligand binds to the RTK, it can induce a conformational change in the receptor, leading to a stable dimerization of the receptor. Dot motif, accompanied by PTK activation. Tyrosine-specific phosphorylation stabilizes the kinase domain in an activated conformation and provides recognition for downstream signal transduction molecules containing the SH2 domain of the src oncogene family and the phosphorylated tyrosine binding domain (PTB) , docking and combining parts. These signal transduction proteins can further recruit some other eff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12Q1/02
Inventor 贺浪冲周亚丽张彦民罗文娟李淼孙萌
Owner XI AN JIAOTONG UNIV
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