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A kind of recombinant hek293 cell with high expression of ephrinb2 and its application

A pez-m61-ephrinb2, recombinant cell technology, applied in the direction of cells modified by introducing foreign genetic material, microorganism assay/inspection, biochemical equipment and methods, etc., can solve the full-length sequence EphrinB2 vector construction and biological application Research and its application have not been reported and other problems, to achieve the effect of improving sensitivity and specificity, and complete molecular activity

Inactive Publication Date: 2018-04-17
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, there are many studies on the EphrinB2 gene, but there are no reports on the construction of the full-length sequence EphrinB2 vector and its biological application research and application.

Method used

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  • A kind of recombinant hek293 cell with high expression of ephrinb2 and its application
  • A kind of recombinant hek293 cell with high expression of ephrinb2 and its application
  • A kind of recombinant hek293 cell with high expression of ephrinb2 and its application

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Embodiment Construction

[0028] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0029] The present invention first constructs the eukaryotic expression vector pEZ-M61-EphrinB2 of the full-length gene of EphrinB2, and its gene sequencing result is consistent with that of human EphrinB2 in the gene bank. Recombinant HEK293 cells that stably and highly express EphrinB2 molecules, that is, recombinant HEK293-EphrinB2 cells. The following detailed description of this aspect is to explain rather than limit the present invention.

[0030] a. the present invention is based on pEZ-M61-EphrinB2 vector (GeneCopoeia company), the primer sequence is as follows:

[0031] Upstream primer: 5-TCATCTTCATCGTCATCATCATC-3

[0032] Downstream primer: 5-CTGTCCGCAGTCCTTAGC-3

[0033] The plasmid map of the constructed pEZ-M61-EphrinB2 vector is as follows figure 1 shown.

[0034] b. HEK293 cells transfected with pEZ-M61-EphrinB2 expression vector

[0035] T...

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Abstract

The invention discloses a recombinant HEK293 cell with high EphrinB2 expression and application thereof. The recombinant cell uses the HEK293 cell as a host cell, and the exogenous expression vector for transfecting the host cell is an expression vector containing a full-length EphrinB2 gene. Compared with the existing recombinant cells constructed only with the EphrinB2 extracellular region or partial gene sequence, the recombinant HEK293 cell membrane constructed by the present invention containing the full length of the EphrinB2 gene can stably express the EphrinB2 molecule and has complete molecular activity. MTT method detected that dasatinib had a good effect on recombinant HEK293 cells at a certain concentration, so it could be used in drug screening targeting EphrinB2.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the construction of non-cancer cells with high expression of EphrinB2, in particular to a recombinant HEK293 cell with high expression of EphrinB2 based on exogenous recombinant plasmid and its application. Background technique [0002] Receptor tyrosine kinases (RTKs) are a huge family that play a very important role in the life activities of organisms. The extracellular region of RTK is the binding ligand domain, and the ligands are soluble or membrane-bound polypeptides or protein hormones, including insulin and various growth factors. The intracellular segment is the catalytic site of the tyrosine protein kinase and has an autophosphorylation site. When the ligand binds to the RTK, it can induce a conformational change in the receptor, leading to a stable dimerization of the receptor, phosphorylation of the dimerized receptor, and formation of a phosphorylated tyrosine site in the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12Q1/02
Inventor 贺浪冲张彦民刘艳萍代秉玲马维娜刘瑞王文捷
Owner XI AN JIAOTONG UNIV
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