Gene engineering preparation method of Chinese human being costimulation signal inhibition factor

A costimulatory signal, inhibitory factor technology, applied in genetic engineering, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as the application of unfavorable genetic engineering products

Inactive Publication Date: 2005-01-12
FUDAN UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no research report on Chinese co-stimulatory signal inhibitors, which is not c...

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
  • Gene engineering preparation method of Chinese human being costimulation signal inhibition factor
  • Gene engineering preparation method of Chinese human being costimulation signal inhibition factor
  • Gene engineering preparation method of Chinese human being costimulation signal inhibition factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] The CTLA-4 gene of Chinese leukocyte DNA was amplified in vitro with artificially synthesized specific primers, and the amplified product was subjected to 1.5% agarose electrophoresis. 3) Precipitate the DNA fragment of the target gene with 80% ethanol, hydrolyze it with the corresponding endonuclease, connect it with the vector to construct the pPICLA plasmid, and store it at 4°C and 0.1M CaCl 2 Transformed KM71 and X33 strains in the presence of presence, cultured in MGYH medium at 30°C until OD 600 =2-6, centrifuge the cells and resuspend them in 100 times AIGYH, culture to OD 600 =2-6, add methanol every day until the final concentration reaches 0.5%, culture for 1-5 days, collect the cells by centrifugation, break the cells, wash with 2M urea, centrifuge to precipitate, then dissolve the precipitate with 8M urea, dialyze with PBS, polyethylene glycol Concentrate, further purify by 8% PAGE (pH8.8) and HPLC, collect the target protein, identify by Western blot, MLR ...

Embodiment 2

[0067] The CTLA-4 gene of Chinese leukocyte DNA was amplified in vitro with artificially synthesized specific primers, and the amplified product was subjected to 1.5% agarose electrophoresis. 3) Precipitate the DNA fragment of the target gene with 80% ethanol, hydrolyze it with the corresponding endonuclease and connect it with the carrier to construct a pPBCTLA plasmid, and store it at 4°C and 0.1M CaCl 2 In case of existence, transform competent DH 5α bacteria, and transform positive bacteria in LB medium at 30°C to OD 600 =0.5, quickly transfer to 42°C and culture for 4 hours, collect the bacteria by centrifugation, destroy the bacteria by conventional methods, separate the inclusion bodies by centrifugation, wash and centrifuge with 2M urea for measurement, then dissolve the precipitate with 8M urea, dialyze with PBS, and concentrate with polyethylene glycol , purified by 8% PAGE (pH8.8) or HPLC, collected the target protein, carried out molecular identification by Western...

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 present invention relates to gene engineering preparation method of Chinese costimulation signal inhibiting factor. It isolate the Chinese costimulation signal inhibiting factor gene and reforms the DNA of Chinese gene codon, adopts host preheminant codon, structures natural protein expression carrier series, fits to express in eukaryote and procaryote, identfication showed that said expression product possesses positive strain of Chines costimulation signal inhibiting factor gene.

Description

technical field [0001] The invention relates to a preparation method of a Chinese co-stimulatory signal inhibitor, ie, a gene engineering expression protein of cytotoxic T lymphocyte-associated antigen 4 and its application. Background technique [0002] Co-stimulatory signal inhibitor is a protein molecule mainly expressed on the surface of mature T cells. It is related to the negative regulation of the immune response that T cells participate in. It is being widely valued internationally. Costimulatory signal inhibitors will soon be used as immune tolerance Drugs are used to treat certain autoimmune diseases and to deal with the rejection of organ transplants. To conduct further functional and application research on co-stimulatory signal inhibitors, it is first necessary to obtain costimulatory signal inhibitor gene data. At present, most of the molecular biological data on costimulatory signal inhibitors are applicable to foreigners, and they are all fusion proteins. Th...

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): A61P37/02A61P37/06C12N15/09C12N15/12C12N15/70C12N15/79C12N15/81C12Q1/68
Inventor 朱乃硕
Owner FUDAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap