Recombinant protein and application thereof

A recombinant protein and protein technology, applied in the field of recombinant protein and its application, can solve the problems of inability to generate and prepare anti-myocardial infarction and vascular leakage drugs, so as to improve cardiac function, inhibit vascular leakage, and inhibit myocardial cell apoptosis. Effect

Inactive Publication Date: 2016-12-21
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the above defects or improvement needs of the prior art, the present invention provides a recombinant protein and its application, the purpose of which is to prepare anti-myocardial infarction and anti-vascular leakage...

Method used

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  • Recombinant protein and application thereof
  • Recombinant protein and application thereof
  • Recombinant protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1A

[0035] Preparation of embodiment 1AGGF1 recombinant protein

[0036] AGGF1 protein expression plasmid pet28-AGGF1 was expressed in Escherichia coli BL21, wherein the sequence of AGGF1 was shown in SEQ NO.1. The expression strain was cultured overnight in 20 ml kanamycin-resistant LB medium. On the next day, add the 20 ml medium to 1 L kanamycin-resistant LB medium, and culture on a shaker at 37° C. for 2.5 hours. Then 1 mM IPTG was added to induce protein expression. IPTG is a highly stable lactose analog that inhibits the lac repressor and induces the synthesis of β-galactosidase. This enzyme promotes the utilization of lactose. IPTG can be used to induce the expression of target genes regulated by the lac operator. After induction by adding IPTG, E. coli continued to grow in the medium for 5.5 hours. Then centrifuge the culture medium, 4000rpm, 4°C, 10mins. If the protein is not to be isolated immediately, store the E. coli pellet at -20°C. The recombinant protein was...

Embodiment 2A

[0037] Example 2 Preparation of AGGF1 Homologous Recombination Protein

[0038] 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, Proteins with 86%, 85%, 84%, 83%, 82%, 81% and 81% homology were prepared. The pet28 protein expression plasmid for obtaining AGGF1 homologous protein was constructed, and prepared according to the full-length AGGF1 protein recombination method, and the preparation method was the same as that in Example 1.

Embodiment 1

[0040] The angiogenesis-promoting effect of the recombinant protein synthesized in Example 1 and its application in the preparation of anti-myocardial infarction drugs

[0041] The C57BL / 6 mice were divided into 3 groups: sham operation group (Sham), placebo group (MI+IgG) for myocardial infarction surgery, recombinant AGGF1 protein group (MI+AGGF1) for myocardial infarction surgery, 12 mice per group Only. Before surgery and after myocardial infarction surgery, the cardiac function of mice was evaluated with a small animal ultrasound imaging system. One day after the operation, the mice were examined by ultrasound to confirm that the myocardial infarction operation was successful. One week after the successful operation, mice were injected with recombinant AGGF1 protein or the same dose of IgG as a control by tail vein, twice a week for 2 weeks. The protein dosage is: 0.25mg / kg.

[0042] Compared the number of new blood vessels (CD31 staining) in the hearts of mice in the ...

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Abstract

The invention discloses a recombinant protein AGGF1. The recombinant protein AGGF1 can improve the cardiac function of a mouse with acute myocardial infarction and inhibit myocardial apoptosis and cardiac fibrosis by promoting angiogenesis in the heart of the mouse. Thus, the recombinant protein AGGF1 can be used for preparation of angiogenesis promoting drugs for treatment of ischemic diseases. Meanwhile, the recombinant protein AGGF1 can improve the cardiac function of the mouse and inhibit cardiac fibrosis by inhibiting vascular leakage and oedema in cardiac muscles after the operation of cardiac ischemia reperfusion. Thus, the recombinant protein AGGF1 provided by the invention can be used for preparation of drugs for inhibiting vascular leakage to treat vascular leakage, oedema and dysfunction caused by cardiac ischemia reperfusion.

Description

technical field [0001] The invention belongs to the field of biomedicine, and more specifically relates to a recombinant protein and its application. Background technique [0002] Acute myocardial infarction (AMI) is a more serious manifestation of coronary atherosclerotic disease (CAD), one of the leading causes of death worldwide. In the United States alone, there is one additional MI patient per year for every 620,000 Americans. 295,000 people have recurrent myocardial infarction, and nearly 400,000 myocardial infarction patients will die because of its sudden onset. In addition, an estimated 150,000 MIs per year go unrecorded. Acute myocardial infarction is caused by coronary artery occlusion. It disturbs the coronary blood flow to the myocardium, and finally leads to myocardial cell necrosis and cardiac function damage. [0003] Recent statistics show that acute myocardial infarction has surpassed valvular heart disease and arterial hypertension [0004] It is the ...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P9/00A61P9/10A61P7/10
CPCC07K14/47A61K38/00
Inventor 王擎姚雨峰吕丘仑
Owner HUAZHONG UNIV OF SCI & TECH
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