Polypeptide, DNA molecule encoding the polypeptide, vector, preparation method and use

a dna molecule and polypeptide technology, applied in the field of biotechnology, can solve the problems of serious consequences of amputation, influence on appearance as well as physical and psychological health, and complex mechanisms, and achieve the effects of no side effects, promotion of tissue repair and wound healing, and promotion of skin tissue repair

Inactive Publication Date: 2016-12-29
WUHAN MORE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0146]The present invention further provides a novel medicament, which is more effective in treating wound surface bacterial infection, acne and eczema. The novel medicament is formulated with an active polypeptide from the traditional Chinese medicinal material scorpion as an active ingredient. The medicament has significant therapeutic effect for the treatment of wound surface bacterial infections, chronic refractory infection wound surfaces on the body surface, acne and eczema, and for the promotion of tissue repair and wound healing. The present invention also rel...

Problems solved by technology

If infection happens, there may leave scars, influencing the appearance as well as physical and psychological health.
Recently, with the intervention of modern cell biology and molecular biology studies, cell activities and influence factors thereof during would healing have been observed in a large number of experiments, and it was considered that various growth factors were involved in the regulation of the wound healing process, with the possible mechanisms being very complex.
Due to features such as development on the body surface, long course of disease, large influences on appearance, high incidence of complications and extremely high treatment expenses, there will be large influences on lives and qualities of patients, which is an important issue that modern society must face.
Due to the lack of clinically effective medicaments for treatment, the serious consequences of amputation may happen.
Recurrence risk is high and is accompan...

Method used

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  • Polypeptide, DNA molecule encoding the polypeptide, vector, preparation method and use
  • Polypeptide, DNA molecule encoding the polypeptide, vector, preparation method and use

Examples

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example 1

Preparation of the Gene for the Broad-Spectrum Antiseptic Active Polypeptide from Chaerilus tryznai Kovarik

[0152]A: Extraction of Total RNA from the Venom Gland of Chaerilus tryznai Kovarik (Trizol LS One-Step Method: Trizol LS was Purchased from Invitrogen, USA)

[0153](1) 500 mg of the venom gland from the scorpion's tail was ground into fine powders in liquid nitrogen, added with 10 mL of TRIZOL reagent with well mixing, and placed at room temperature (20-25° C., the same as hereinafter) for 5 min. (2) 2 mL chloroform was then added with mixing for 15 s, placed at room temperature for 2-3 min, and centrifuged at 12000 g and 4° C. for 15 min. (3) The aqueous phase was taken out and added with same volume of isopropanol, then placed at room temperature for 10 min, and centrifuged at 12000 g and 4° C. for 10 min to obtain RNA precipitate. (4) The precipitate was washed with 5 ml of 75% ethanol and centrifuged at 7500 g for 5 min. (5) The RNA precipitate was dried followed by dissolut...

example 2

Structure Analysis of Ctryamp Polypeptide and the Structurally Homologous Amphiphilic Polypeptides Thereof

[0170]According to the sequence (FIRIARLLRIF) of the mature peptide of Ctryamp as set forth in SEQ ID NO: 2 as provided in EXAMPLE 1, the secondary structure of Ctryamp was predicted by using on-line NPS@ server [DSC method (Discrimination of protein Secondary structure Class)], and shown by software AHTHEPROT 2000. Results showed that, the Ctryamp contained 100% of α-Helix structure, had a typical amphiphilic α-Helix structure and comprised a large number of basic residues (Arg) with net positive charges. Based on the helix diagram of the polypeptide sequence, a large number of point mutations on the polypeptide sequence FIRIARLLRIF of Ctryamp were performed then. It was found that the sequence FIX1IAX2LLX3IF (X1, X2 and X3 independently are any one of three basic amino acids His, Arg and Lys) (SEQ ID NO: 1) did not influence its amphiphilic property (Table 1). Therefore, the p...

example 3

Polypeptide and the Structurally Homologous Amphiphilic Polypeptides Thereof

[0171]Artificial synthesis was carried out according to the amino acid sequence of the Ctryamp (FIRIARLLRIF) as provided in EXAMPLE 1 and that of the structurally homologous amphiphilic polypeptides thereof (FIX1IAX2LLX3IF, as provided in EXAMPLE 2). A high-purity Ctryamp polypeptide (as shown in FIGS. 2 and 3) and structurally homologous amphiphilic polypeptides thereof were obtained by solid-phase chemical synthesis (Table 1).

Example 4

Bacteriostasis Experiment of Ctryamp Polypeptide and the Structurally Homologous Amphiphilic Polypeptides Thereof

[0172]Bacteriostasis Experiment on Gram-Negative Bacteria:

[0173]96-well plate culturing method: (1) When Pseudomonas aeruginosa (including a standard strain, clinically isolated strain and drug resistant strain), Escherichia coli (including standard strains CCTCC AB94012 and ATCC25922, a clinically isolated strain and drug resistant strain), Klebsiella pneumoniae, ...

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Abstract

A polypeptide, a DNA molecule encoding the polypeptide, a vector, a preparation method and a use therefor are disclosed. The polypeptide comprises an amino acid sequence represented by formula (I) or formula (II): formula (I) comprises an amino acid sequence represented by SEQ ID NO: 1; formula (II) comprises an amino acid sequence obtained by subjecting the amino acid sequence represented by SEQ ID NO: 1 to modification, substituted, and deletion or addition of one or more amino acids. The polypeptide may be used in the preparation of drugs that treat or prevent diseases related to infections caused by bacteria, and can also be used in the preparation of drugs that promote tissue repair and wound healing.

Description

[0001]The present application claims the priorities of Chinese Patent Application No. 201310300278.6, entitled “POLYPEPTIDE, DNA MOLECULE ENCODING THE POLYPEPTIDE, VECTOR, PREPARATION METHOD AND USE” filed with the Chinese State Intellectual Property Office on Jul. 17, 2013, and Chinese Patent Application No. 201410327573.5, entitled “POLYPEPTIDE, DNA MOLECULE ENCODING THE POLYPEPTIDE, VECTOR, PREPARATION METHOD AND USE” filed with the Chinese State Intellectual Property Office on Jul. 10, 2014, which are both incorporated herein by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention relates to the field of biotechnology, particularly to a polypeptide, DNA molecule encoding the polypeptide, vector, preparation method and use.BACKGROUND OF THE INVENTION[0003]Among various wounds, no matter in the clinical treatment of various injuries such as burn injury, cold injury, crush injury, war injury, animal bite, nuclear radiation injury and combined injury, or in ...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K7/06A23K20/195A61K38/08
CPCC07K7/08A61K38/00C07K7/06A61K38/08A61K38/10C12N15/63A61P31/04A61P17/02Y02A50/30
Inventor LI, WENXINCAO, ZHIJIANWU, YINGLIANGWANG, ZESHENGHAN, SONG
Owner WUHAN MORE BIOTECH
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