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Cell factor FAM19A4 with anti-infection and antineoplastic activity and application thereof

A host cell and antibody technology, applied in the direction of anti-cytokine/lymphokine/interferon immunoglobulin, cytokine/lymphokine/interferon, anti-tumor drugs, etc., can solve the problem of increasing the probability of infection in immunocompromised people

Inactive Publication Date: 2013-11-06
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] On the other hand, with the widespread application of antibiotics, corticosteroid hormones, and immunosuppressants, clinically drug-resistant strains are gradually increasing, and various intracellular and extracellular microbial infections are on the rise, which makes the infection rate of immunocompromised people Substantial growth

Method used

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  • Cell factor FAM19A4 with anti-infection and antineoplastic activity and application thereof
  • Cell factor FAM19A4 with anti-infection and antineoplastic activity and application thereof
  • Cell factor FAM19A4 with anti-infection and antineoplastic activity and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0084] Embodiment 1, cDNA cloning and bioinformatics analysis of FAM19A4 gene

[0085] 1. Bioinformatics analysis

[0086] The cDNA sequence and alternative splicing bodies were analyzed by comparing with NCBI's Human est database. Homologous proteins were searched through NCBI's tBLASTn program, and at the same time from the genome database ( http: / / www.ncbi.nlm.nih.gov / genome / ) to get the chromosome location. Protein functional domain analysis using NCBI CDD database ( http: / / www.ncbi.nlm.nih.gov / Structure / cdd / cdd.shtml ). Other protein-related analyzes used the ExPASy database ( http: / / us.expasy.org / ), including: TMHMM ( http: / / www.cbs.dtu.dk / services / TMHMM / ) for transmembrane region analysis; Signal P( http: / / www.cbs.dtu.dk / services / SignalP / ) for signal peptide analysis; Psort II ( http: / / psort.nibb.ac.jp ) for subcellular localization prediction; GNF SymAtlas (http: / / symatlas.gnf.org / SymAtlas / ) for expression profile analysis; Prosite ( http: / / www.expas...

Embodiment 2

[0100] Example 2, FAM19A4 expression profile analysis

[0101] The human normal tissue cDNA library purchased from clontech and the human cDNA library prepared as described above were used to analyze the mRNA expression level of FAM19A4 in normal tissues and leukemia cell lines. The specific operation is as follows:

[0102]FAM19A4 was amplified using the PCR amplification conditions described in Example 1-2. For the mouse cDNA library prepared in 1 above, use 5' primer (5'-TTCACGGTCACTCTCCAGGAC-3', SEQ ID No.5) and 3' primer (5'-AGCCACGTAAATGCCACCTTC-3', SEQ ID No.6) to amplify To increase the mouse homologous gene Fam19a4, the amplification conditions are 94°C (5 minutes) → 94°C (40 seconds), 54°C (40 seconds), 72°C (40 seconds), 30 cycles of amplification → 72°C (7 minute).

[0103] Use 5' primer (5'-TGAAGGTCGGAGTCAACGGATTTGGT-3', SEQ ID No.7) and 3' primer (5'-CATGTGGGCCATGAGGTCCACCAC-3', SEQ ID No.8) to amplify GAPDH as an internal reference, and the amplification cond...

Embodiment 3

[0109] Example 3, Construction of FAM19A4 Eukaryotic Expression Plasmid and Purification of Eukaryotic Protein

[0110] 1. Construction of FAM19A4 eukaryotic expression plasmid

[0111] In order to carry out the functional research of FAM19A4, this example uses Not I and Kpn I to digest the pGEM-T-FAM19A4 recombinant plasmid, recovers and further ligates the target gene fragment (coding region of FAM19A4) into The eukaryotic expression vector pcDNA3.1-myc / his(-)B (Invitrogen, V85520) was used to obtain the recombinant eukaryotic expression plasmid of pcDB-FAM19A4-myc-his (fused with c-myc and his tags).

[0112] 2. Expression and purification of recombinant FAM19A4 protein

[0113] 293T cells were plated in 10 cm culture dish with DMEM medium containing 10% FBS in 5% CO 2 , and cultivated in a 37°C incubator for 24 hours. The target gene pcDB-FAM19A4-myc-his eukaryotic expression plasmid was transfected by Vigofect cationic transfection method. After 24 hours of transfecti...

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Abstract

The invention relates to a cell factor FAM19A4 with anti-infection and antineoplastic activity and application thereof. Specifically speaking, the invention provides a gene or protein of FAM19A4 or immunological fragments thereof, genetic engineering vectors and host cells including the gene or protein of FAM19A4 or the immunological fragments thereof and antibodies to the protein of FAM19A4 or the immunological fragment thereof. The invention further provides application of the gene or protein of FAM19A4 or the immunological fragments thereof in preparation of medicinal preparations having antiviral, anti-endocellular bacterial, antimycotic, anti-extracellular bacterial and antineoplastic effects. Furthermore, the invention also provides application of a detection reagent in preparation of compositions used for auxiliary diagnosis and prognosis of diseases related to immunization.

Description

technical field [0001] The present invention relates to a new human cytokine and its application, in particular to the gene or protein of FAM19A4 or their immune fragments, and the gene or protein of FAM19A4 or their immune fragments have anti-infection and anti-tumor activities. Background technique [0002] Cytokines are small molecular proteins secreted by various cells in the body that regulate cell growth and differentiation, regulate immune function and physiological responses, and participate in pathological responses, and play a role by binding to receptors. Cytokines mainly include interleukin (Interleukin, IL), colony-stimulating factor (Colony-Stimulating Factor, CSF), interferon (Interferon, IFN), tumor necrosis factor (Tumor-Necrosis Factor, TNF), chemokine (Chemokine )Wait. Cytokines are the language of communication between immune cells and play an important role in both innate and adaptive immune responses. [0003] The innate immune response is the body's ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/52C07K16/24C12N15/19C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12Q1/68G01N33/53A61K38/19A61K39/395A61K48/00A61P35/00A61P31/04A61P31/10A61P31/12A61P37/02
Inventor 韩文玲马大龙王文彦张荷钰王平章张颖妹宋泉声李婷付伟伟潘文
Owner PEKING UNIV
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