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Secretion-type FNDC5 protein and preparation method and application thereof

A secreted, protein technology, applied in the field of genetic engineering, to achieve the effect of convenient access, improvement of glucose and insulin tolerance, and improvement of insulin resistance

Active Publication Date: 2020-01-14
THE SECOND HOSPITAL OF SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Directly secreted FNDC5 isoforms without a transmembrane domain and their use in lipid metabolism have not been reported so far

Method used

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  • Secretion-type FNDC5 protein and preparation method and application thereof
  • Secretion-type FNDC5 protein and preparation method and application thereof
  • Secretion-type FNDC5 protein and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0026] A secreted FNDC5 amino acid sequence is shown in SEQ ID NO: 1;

[0027] MPPGPCPRAALRLWLGCVCFALVQADAPVNVTVRHLKANSAVVSWDVLEDEGFAISQQKKDVRMLRFIQEVNTTTRSCALWDLEEDTEYIVHVQAISIQGQSPASEPVLFKTPREAEKMASKNEVPFANVDDTGKHNQRAALQNAEGAKA;

[0028] Preparation method of secreted FNDC5 protein:

[0029] (1) Design primers from different sites of the fndc5 gene, perform PCR, run electrophoresis, and detect other isomers besides the known fndc5 isomers; specifically,

[0030] Primers are:

[0031] Upstream primer FNDC5-F: TGAGGCCGAGAAGATGGCCTCTAA;

[0032] Downstream primer FNDC5-R: TAGTGACAATGGCTGCTCTCTGCC;

[0033] Extract INS-1 cell RNA, reverse transcribe into cDNA, and use the above primers for PCR.

[0034] PCR reaction system: H 2 O:36.75ul

[0035] Go Taq DNA Polymerase: 0.25ul (Promega)

[0036] 5x Go Taq Buffer: 10ul (Promega)

[0037] FNDC5-F: 1ul

[0038] FNDC5-R: 1ul

[0039] cDNA: 1ul

[0040] Total 50ul

[0041] 94°C for 5 minutes;

[0042] 94°C for 30 seconds,...

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Abstract

The invention discloses a secretion-type FNDC5 protein. The secretion-type FNDC5 protein does not contain a transmembrane domain and can be directly secreted and released into blood; and the amino acid sequence of the FNDC5 protein is SEQ ID NO:1. The secretion-type FNDC5 protein is simple in structure and convenient to obtain. The secretion-type FNDC5 protein can be used for treating obesity-related metabolic disorders; after the secretion-type FNDC5 recombinant protein is injected into the abdominal cavity of a high-fat fed obese mouse model, glucose and insulin tolerance can be effectivelyimproved, and it shows that the protein can obviously relieve glucose metabolic disorders and insulin resistance of obese patients; and fatty liver occurrence can be obviously relieved, the level of plasma cholesterol, free fatty acid and triglyceride is decreased, and it shows that the protein can relieve the lipometabolic disorders of the obese patients. The secretion-type FNDC5 protein can serve as a new drug target for treating the metabolic disease to be developed.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a secreted FNDC5 protein and its preparation method and application. Background technique [0002] Obesity is a common metabolic disease that threatens the health of people all over the world. Obese patients are more likely to develop cardiovascular and cerebrovascular diseases, type 2 diabetes, respiratory distress syndrome and musculoskeletal diseases. The energy intake of the human body is higher than the energy consumption, and the remaining calories will be stored in the body in the form of adipose tissue. When the content of adipose tissue exceeds the physiological needs, it will evolve into obesity. Therefore, increasing energy expenditure has gradually become a powerful means for the potential prevention and treatment of obesity and its related diseases. [0003] Irisin, a protein with 112 amino acid residues discovered in 2012, is the product of cleavage of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/575C12N15/81A61K38/22A61P3/04C12R1/84
CPCA61K38/00A61P3/04C07K14/575C12N15/815
Inventor 唐东起李世武李慧
Owner THE SECOND HOSPITAL OF SHANDONG UNIV
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