Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Protein AGGF1 and application of FHA polypeptide of protein AGGF1 in preparation of anti-inflammatory drug

An anti-inflammatory drug and pharmaceutical technology, applied in the field of biomedicine, can solve the problem that anti-inflammatory drugs can only specifically treat inflammatory diseases, achieve ICAM and chemokine IL-8 expression inhibition, inhibit the occurrence of vascular inflammation, reduce The effect of adhesion factor E-Selectin

Active Publication Date: 2013-08-21
田小利
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the problem that the existing anti-inflammatory drugs can only specifically treat inflammatory diseases, the embodiment of the present invention provides a new application of the protein AGGF1 and its FHA polypeptide, specifically the application in the preparation of anti-inflammatory drugs

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Protein AGGF1 and application of FHA polypeptide of protein AGGF1 in preparation of anti-inflammatory drug
  • Protein AGGF1 and application of FHA polypeptide of protein AGGF1 in preparation of anti-inflammatory drug
  • Protein AGGF1 and application of FHA polypeptide of protein AGGF1 in preparation of anti-inflammatory drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Obtaining the nucleotide sequences of AGGF1, dFHA-AGGF1, FHA.

[0037] Methods: Using pcDNA3.1-AGGF1 plasmid as a template, the nucleotide sequence of AGGF1, d FHA-AGGF1, FHA was amplified by PCR.

[0038] Material: pcDNA3.1-AGGF1 plasmid (see Tian XL, Kadaba R, You SA, Liu M, Timur AA, Yang L, Chen Q, Szafranski P, Rao S, Wu L, Housman DE, DiCorleto PE, Driscoll DJ for the method of obtaining , Borrow J, Wang Q. Identification of an angiogenic factor that when mutated causes susceptibility to Klippel-Trenaunay syndrome. Nature. 2004Feb12; 427(6975): 640-645.), conventional PCR reagents, forward primer and reverse primer ( Synthesized by Beijing Aoke Dingsheng Biotechnology Co., Ltd.), PCR product purification kit (provided by Bomide).

[0039] step:

[0040] 1. PCR reaction (polymerase chain reaction)

[0041] 1) Primers include:

[0042] AGGF1-subclone-F: as shown in SEQ ID NO:4;

[0043] AGGF1-subclone-R: as shown in SEQ ID NO:5;

[0044] dFHA-AGGF1-...

Embodiment 2

[0075] Embodiment two: prepare the plasmid vector containing the nucleotide sequence gained in embodiment one

[0076] Method: The nucleotide sequences of AGGF1, dFHA-AGGF1 and FHA obtained in Example 1 were respectively inserted into the p-shuttle-IRES-GFP plasmid vector to obtain a plasmid vector containing the nucleotide sequence of AGGF1, containing dFHA-AGGF1 core The plasmid vector of the nucleotide sequence, the plasmid vector containing the FHA nucleotide sequence.

[0077] Materials: p-shuttle-IRES-GFP plasmid (provided by stratagene), restriction enzymes, T4DNA ligase (provided by NEB Company), DH5a competent cells (provided by Tiangen Biochemical (Beijing) Technology Co., Ltd.) , Plasmid extraction kit (provided by Bomed).

[0078] step:

[0079]1. The three PCR products obtained in Example 1 were digested with the p-shuttle-IRES-GFP plasmid for 6 hours, and the digested products of the PCR products and p-shuttle-IRES-GFP were recovered respectively using the PCR...

Embodiment 3

[0088] Example 3: Construction and purification of adenoviral vector

[0089] Objective: To construct an adenovirus vector containing the plasmid vector obtained in Example 2, and to amplify and purify it.

[0090] Materials: p-shuttle-AGGF1, p-shuttle-d FHA-AGGF1, p-shuttle-FHA, HEK293A (purchased from ATCC), HUVEC cells (purchased from ATCC), CsCl (1.4g / ml and 1.2g / ml) , PmeI endonuclease (provided by NEB Company), PacI endonuclease (provided by NEB Company), BJ5183 competent bacteria (provided by addgene Company), virus purification column (GE healthcare Capto TM DeVirS. Capto DeVirS).

[0091] step:

[0092] 1. Linearize p-shuttle-AGGF1, p-shuttle-d FHA-AGGF1, and p-shuttle-FHA with PmeI, and recover the digested fragments;

[0093] 2.shuttle plasmid and pAdeasy-1Vector (by Provided by Stratagene ) to co-electroporate BJ5183 competent bacteria;

[0094] 3. Extract the kanamycin-positive plasmid, digest it with PacI, and transfect HEK293A cells;

[0095] Transfection...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a novel protein AGGF1 with high anti-inflammatory performance, FHA polypeptide of the protein AGGF1 and application of the FHA polypeptide in preparation of an anti-inflammatory drug, and belongs to the field of biological medicine. The FHA polypeptide represented by SEQ ID NO:1 in a sequence list and the protein AGGF1 represented by SEQ ID NO:2 in the sequence list are expressed through cloning; and effects of the FHA polypeptide and the protein AGGF1 in an endothelial cell HUVEC and mononuclear cell U937 adhesion process, effects of the FHA polypeptide and the protein AGGF1 for expressing inflammatory factors E-Selectin, ICAM and IL-8 in the endothelial cells HUVEC and effects of the FHA polypeptide and the protein AGGF1 for inhibiting the activity of an NF-kB starter are checked, so that the FHA polypeptide and the protein AGGF1 can be found to be capable of inhibiting the activation of the blood vessel endothelium and then further inhibiting the generation of vascular inflammation. Therefore, the polypeptide comprised in an amino acid sequence provided by the invention or consisting of the SEQ ID NO:1 or the SEQ ID NO:2 can be effectively used for preparing the anti-inflammatory drug and lays the foundation for the preparation of broad-spectrum anti-inflammatory drugs and the prevention or treatment of inflammation related diseases.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of a protein AGGF1 and its FHA polypeptide in the preparation of anti-inflammatory drugs. Background technique [0002] Inflammation refers to the defense response of living tissue with vascular system to injury factors, and the response of inflammation to the damaged local tissue of the body is called inflammatory response. Vascular inflammatory response is the central link of inflammatory response. Inflammation can lead to the occurrence of various human diseases, such as cardiovascular disease, neurodegenerative disease, metabolic disease, tumor and aging. In the development of inflammatory diseases, the vascular inflammatory response is an inevitable response to the inflammatory response. Vascular inflammation can lead to cardiovascular diseases such as atherosclerosis and acute coronary syndrome, seriously affecting people's quality of life, and even bringing lif...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/00A61K38/16C12N15/11C12N15/63C12N5/10C12N7/01C12N5/078A61P31/00
Inventor 田小利胡方园张勇
Owner 田小利
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com