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Interleukin-17 acceptor, and its coding gene and application

A technology of interleukins and receptors, applied in the fields of proteins and their coding genes and applications, to achieve great practical significance and broad application prospects

Inactive Publication Date: 2008-09-10
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current understanding of IL-17 ligand-receptor interaction is still in its infancy, and many issues still need to be further studied

Method used

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  • Interleukin-17 acceptor, and its coding gene and application
  • Interleukin-17 acceptor, and its coding gene and application
  • Interleukin-17 acceptor, and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, the cloning of mIL-17RE

[0033] Extract the total RNA of mouse colon tissue according to the conventional method and use it as a template under the guidance of primer P1 (upstream primer): 5′-cagacaggaaagacatggtctc-3′ and P2 (downstream primer): 5′-agacagctgaaaccacagggac-3′ The one-step RT-PCR kit from TaKaRa Company was used for RT-PCR amplification according to the instructions of the kit. After the reaction, the PCR amplification product was detected by 1% agarose gel electrophoresis, and the target fragment of 1914bp was recovered and purified. Since an A base was added to the 5' end of the product amplified by Taq polymerase, the target fragment was directly subcloned into the vector pGEM-T (purchased from Promega), and the recombinant product was transformed into Escherichia coli DH5α. State cells, positive clones were obtained by blue-white screening, the positive clones were selected to extract plasmids, and the plasmid vector containing the targ...

Embodiment 2

[0034] Example 2, bioinformatics analysis of mIL-17RE

[0035] Bioinformatics analysis is carried out to the amino acid residue sequence (SEQ ID №: 2 in the sequence listing) of the encoded protein mIL-17RE of mIL-17RE, and the analysis results are as follows: figure 2 As shown, the 1st to 25th amino acid residues from the amino terminal (N-terminal) of mIL-17RE1 is the signal peptide sequence, which consists of 25 amino acid residues; the 26th to 413th amino acid residues from the N-terminal are the extracellular region The sequence consists of 388 amino acid residues; the 414th to 439th leucine from the N-terminal is the transmembrane region sequence, which consists of 26 amino acid residues; the 440th to 637th leucine from the N-terminal is the intracellular region sequence, consisting of 198 amino acid residues. The results of ProSite analysis showed that there were two typical N-glycosylation sites in the extracellular region of mIL-17RE1, which were located at the 278t...

Embodiment 3

[0036] Embodiment 3, Northern hybridization detects the expression situation of mIL-17RE gene

[0037] The expression of mIL-17RE gene in normal mouse tissues and embryos was detected by Northern hybridization. The specific method is as follows: 8-week-old normal Kunming mice were killed by neck dislocation, and the main organs were collected, including heart, liver, spleen, kidney, lung, brain, stomach and testis. And after the mice were mated, embryos with gestational ages of 7.5, 8.5, 9.5, 10.5, 11.5, 12.5, 13.5 and 14.5 days were taken respectively. After the above organs and embryos were ground and homogenized, they were treated with TRIzol (purchased from Invitrogen) to extract total tissue RNA. About 10 μg of total RNA from each tissue was separated on 1.3% formaldehyde gel, and then transferred to a membrane of Hybond (purchased from Amersham Biosciences). Using the full-length cDNA of mIL-17RE as a template, the Prime-A-gene kit (purchased from Promega) was used to ...

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Abstract

This invention discloses an acceptor of inferleukin 17 and its code and their application, the objective is to provide an acceptor of inferleukin 17 and its code gene and their application of preparing mark substance for diagnose tumour and treatment medicine for RAS / MAPK signal access related diseases. The albumen is a protein with following one of the sequence of amino-acid residue radical: 1) SEQ ID No: 2 in sequence table; 2) amino-acid residue radical sequence in SEQ ID No: 2 in the sequence table are replaced by one to ten of amino-acid residue radical, lose or added and through activating RAS / MAPK signal access and with promoting karyokinesis action protein. inferleukin 17 acceptorof this invention will play a important roll in diagnose tumor, since its code gene mIL-17RE interferes the code gene of RNA, it can be used as active component to prepare medicine, for treatment diseases related to RAS / MAPK signal access, such as prostate tumour, leukaemia and thyroid gland tumour and other diseases. The invention has more practical meaning and wide application prospect in medical science and biological pharmacy field.

Description

technical field [0001] The present invention relates to proteins and their coding genes and applications, in particular to a mouse-derived interleukin 17 receptor and its coding genes and their use in the preparation of tumor diagnostic markers and therapeutic drugs for diseases related to RAS / MAPK signaling pathways Applications. Background technique [0002] The interleukin 17 (IL-17) and its receptor (IL-17R) family is a unique immune molecule-receptor family that plays an important role in normal body immunity, inflammatory response and tumorigenesis (J.K.Kolls, A . Linden, Interleukin-17 family members and inflammation. Immunity 21(4) (2004) 467; T.A. Moseley, D.R. Haudenschild, L. Rose, A.H. Reddi, Interleukin-17 family and IL-17 receptors. Cytokine Growth Factor Rev 14(2 ) (2003) 155). Currently known IL-17 ligands have six members, and each ligand acts on a different immune process. For example, the earliest discovered IL-17A can stimulate a variety of tissue cell...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/705C07H21/04C12N15/24C12N5/10G01N33/68
Inventor 刘力李铁石李雪妮傅新元常智杰张淑平
Owner TSINGHUA UNIV
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