Secretory protein with migration activity and application thereof

A protein and homology technology, applied in the field of genetic engineering, can solve the problems of large investment and heavy workload, and achieve the effect of broad application prospects

Active Publication Date: 2012-11-14
PEKING UNIV
View PDF1 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These tasks will take far more time, greater investment and heavier

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
  • Secretory protein with migration activity and application thereof
  • Secretory protein with migration activity and application thereof
  • Secretory protein with migration activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Embodiment 1, construction pcDNA3.1-FAM19A5-myc-his6 fusion protein expression plasmid

[0071] The pcDNA3.1-FAM19A5-myc-his6 plasmid was constructed to express the FAM19A5-myc-his6 fusion protein.

[0072] 1. Method:

[0073] The FAM19A5 coding sequence (SEQ ID NO: 2; 3; 4) was respectively inserted into the pcDNA3.1-myc-his6 (Invitrogen Company) expression vector to construct the pcDNA3.1-FAM19A5-myc-his6 expression plasmid. After the correctness of the plasmid was verified by sequencing, the plasmid was amplified, and the plasmid was extracted with the Axygen plasmid extraction kit for cell transfection.

[0074] 2. Results:

[0075] The sequence of the coding region was correct by DNA sequencing.

Embodiment 2

[0076] Example 2, plasmid transfection cells to obtain FAM19A5 expression supernatant

[0077] 1. Method:

[0078] HEK 293T cells adjusted to 6×10 5 Cells / 2ml concentration in 6-well plate at 37°C 5% CO 2 Culture for 24 hours for transfection, pcDNA3.1-FAM19A5 (BC039396.1; or NM_001082967.1; or NM_015381.5)-myc-his6DNA 2μg, vigofect (Vigofect Biotechnology Co., Ltd.) 2μl, the mixture was dripped slowly Into the prepared cells, at the same time set pcDNA3.1-myc-his6 empty vector transfection cells as a control, change the serum-free medium HEKG after 6 hours, culture at 37°C for 48 hours, and harvest the supernatant after transfection.

[0079] Use Western Blotting to check the expression of the target protein:

[0080] After 12.5% ​​SDS-PAGE electrophoresis, the protein was transferred to the nitrocellulose membrane with Tris-glycine electrotransfer solution, after adding Anti-c-myc antibody (MBL company) to act, then adding IRDyeTM 800-labeled anti-mouse IgG (LICOR Bioscie...

Embodiment 3

[0084] Example 3, the separation of human peripheral blood mononuclear cells

[0085] 1. Method:

[0086] Concentrated white blood cells were provided by Beijing Blood Center, and human peripheral blood mononuclear cells were obtained using lymphocyte separation medium (Shanghai Huajing Bio-Technology Co., Ltd.). Culture with RPMI 1640 (Life Technologies, Inc.) containing 10% heat-inactivated fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin.

[0087] 2. Results:

[0088] Human peripheral blood mononuclear cells were obtained and cultured in vitro for future use.

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 provides a chemotactic leucocyte and an FAM19A5 secretory protein of amino acid sequence as shown in SEQ ID NO: 1 of tumor, a polynucleotide coding the protein, and a polynucleotide-containing genetic engineering carrier. The invention further relates to a drug composite, the protein or polynucleotide-containing genetic engineering carrier and/or host cell, and a pharmaceutically acceptable salt, carrier or excipient. The invention further relates to a method for detecting the expression level change of the protein or the polynucleotide in vitro. Furthermore, the protein or the polynucleotide can be used for preventing and/or curing the virus infection, the allergic diseases, the inflammatory response, the transplant rejection, the cerebral disease, the autoimmunity disease, the tumor metastasis, the immune adjustment, the proliferation and differentiation of stem cells, the osteoporosis, the obesity, the insulin resistance and the cardiovascular disease such as the arteriosclerosis, the angiosteosis and the like, and can be used for commercialized agent to research the symptoms. The protein or the polynucleotide can be further taken as a target to develop the compound, the antibody, the polypeptide drug and the commercialized reagent.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular, the present invention relates to proteins with multiple functions and polynucleotides encoding them, genetic engineering vectors containing polynucleotides, and corresponding pharmaceutical compositions, and the present invention also relates to the proteins And the application of polynucleotide and pharmaceutical composition thereof. Background technique [0002] Cytokines are small molecular soluble proteins that are synthesized and secreted by various cells in the body and have various physiological activities and participate in pathological reactions. Chemokines are a class of cytokines with chemotactic effects, which play a role in the body's viral infection, allergic disease, inflammatory response, transplant rejection, brain disease, autoimmune disease, tumor metastasis, immune regulation, stem cell proliferation and differentiation, bone It plays an important rol...

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): C07K14/52C12N15/19C12N15/63A61K38/19A61K48/00A61P31/12A61P37/08A61P29/00A61P37/06A61P25/00A61P37/02A61P35/04A61P19/10A61P3/04A61P3/10A61P9/10A61P9/14A61P9/12A61P9/04G01N33/68C12Q1/68C07K16/24
Inventor 王应马大龙张焱郑奕郭嫦媛张文娟张颖妹王平章郭帅石太平
Owner PEKING 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