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Wild type EGFR high expressing recombinant HEK 293 cells

A HEK293-EGFR, recombinant cell technology, applied in the direction of cells modified by introducing foreign genetic material, recombinant DNA technology, microbial assay/inspection, etc., can solve the application of wild-type full-length EGFR gene cells related biological characteristics research Issues that have not been reported

Inactive Publication Date: 2011-07-27
XI AN JIAOTONG UNIV
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Problems solved by technology

However, the relevant biological characteristics and applications of wild-type full-length EGFR gene cells have not been reported.

Method used

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  • Wild type EGFR high expressing recombinant HEK 293 cells
  • Wild type EGFR high expressing recombinant HEK 293 cells
  • Wild type EGFR high expressing recombinant HEK 293 cells

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Embodiment Construction

[0029] In the present invention, on the basis of constructing eukaryotic expression carrying pcDNA3.1(+)-EGFR of the wild-type EGFR full-length gene, HEK293 cells are transfected to obtain a recombinant HEK293-EGFR cell strain that stably and highly expresses EGFR molecules.

[0030] Compared with the negative control cells not transfected with the expression vector, the EGFR expression of the recombinant HEK293-EGFR cell line increased by 4 to 5 times, which can be used in drug screening targeting EGFR to increase the sensitivity and specificity of drug screening;

[0031] And compare the wild-type EGFR high expression cell line HEK293-EGFR and the cancerous EGFR high expression cell line A431, apply to the study of the biological characteristics of EGFR and the difference comparison of EGFR pathway proteins, and improve the effectiveness of discovering functional proteins.

[0032] The following is a detailed description of this aspect:

[0033] The present invention is base...

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Abstract

The invention discloses wild type EGFR stably and high expressing recombinant HEK 293-EGFR cells constructed on the basis of exogenous recombinant plasmids pcDNA3.1(+)-EGFR, which can improve the expression of EGFR by 4 to 5 times compared with the expression before transfection. EGF serving as an EGFR accelerator and gefitinib serving as a tyrosine kinase inhibitor are used to induce transfectedHEK293 cells, and the examination of the change of the number of expressed phosphorylated EGFR and ERK in the cells shows that the transfected EGFR has activities; the medicinal gefitinib also has aninhibiting effect on the transfected EGFR, so EGFR-targeted medicament screening can improve the screening specificity and flexibility; and the wild type EGFR stably and high expressing recombinant HEK 293-EGFR cells can be used in comparative researches on the differences of EGFR pathway proteins.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the construction of non-cancer cells with high wild-type EGFR expression, in particular to a recombinant HEK293 cell with high wild-type EGFR expression based on exogenous recombinant plasmids, and its application. technical background [0002] Epidermal growth factor receptor (EGFR) is the expression product of proto-oncogene cerbB1 (HER1), which is highly expressed in many malignant tumors. EGFR is a transmembrane glycoprotein composed of 1186 amino acids with a relative molecular weight of 170kD, and consists of three parts: the extracellular region, the transmembrane region and the intracellular region. The glycosylated extracellular region is folded into 4 subregions, consisting of a ligand binding site and 2 cysteine-rich regions, which can bind a variety of ligands with agonistic functions, mainly including epidermal growth factor, Transforming growth factor α, B cell growth fa...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/79C12N5/10C12Q1/02
Inventor 贺浪冲罗文娟李旭许雪梅王嗣岑葛晓雯李聪
Owner XI AN JIAOTONG UNIV
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