CN103804482A - Antibacterial peptide and application thereof - Google Patents
Antibacterial peptide and application thereof Download PDFInfo
- Publication number
- CN103804482A CN103804482A CN201410067697.4A CN201410067697A CN103804482A CN 103804482 A CN103804482 A CN 103804482A CN 201410067697 A CN201410067697 A CN 201410067697A CN 103804482 A CN103804482 A CN 103804482A
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- Prior art keywords
- antibacterial peptide
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- fish
- antibacterial
- peptide
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/461—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from fish
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to the field of molecular biology and particularly relates to an antibacterial peptide and an application thereof. The antibacterial peptide is represented by an amino acid sequence in the sequence table SEQ ID No.1. The antibacterial peptide is used for preparing an inhibitor for resisting bacterial or virus infection of fishes and the antivirus and antibacterial capabilities of the fishes can be obviously enhanced.
Description
Technical field
The present invention relates to biology field, specifically a kind of antibacterial peptide and application thereof.
Background technology
Antibacterial peptide (antimicrobial peptide) is the positively charged ion small peptide of a kind of tool broad-spectrum antimicrobial, antiviral even Antiparasitic Activity.Antibacterial peptide is incorporated on cytolemma by its distinctive amphipathic molecule structure, thereby destroys cytolemma, kills bacterium.Be different from traditional antibiotic medicine, the bactericidal mechanism of this uniqueness of antibacterial peptide can not make target bacteria produce resistance, and most of antibacterial peptide has the advantages that thermostability is high, so antibacterial peptide more and more causes people's attention.Antibacterial peptide is extensively present in microorganism and animal and plant body, and the antibacterial peptide of having found at present has kind more than 1800.So far the antibacterial peptide identifying in fish comprises hepcidin, defensins, pleurocidin, piscidin, moronecidin, misgurain, pardaxin, parasin, tens of kinds of LEAP-2 and NK-lysin etc.Detect and find, the anti-microbial activity of these antibacterial peptide tool wide spectrums, some also has antiviral activity.Antibacterial peptide is the important component part of fish non-specific immunity system, resists in pathogen infection process and plays an important role fish.Therefore, antibacterial peptide has huge application potential and good application prospect in disease control of aquatic animal.
Summary of the invention
The object of the invention is to provide a kind of antibacterial peptide and application thereof.
For achieving the above object, the technical solution used in the present invention is:
A kind of antibacterial peptide, antibacterial peptide is shown in the aminoacid sequence in sequence table SEQ ID No.1.
The application of described antibacterial peptide, described antibacterial peptide is for the preparation of the inhibitor of antibacterium or virus infection.
Tool of the present invention has the following advantages: antibacterial peptide of the present invention can obviously improve the antiviral and bacterial-infection resisting ability of fish.
Embodiment
Below in conjunction with embodiment, the invention will be further described.Embodiment is intended to the description of giving an example to the present invention, but not limits the invention in any form.
Embodiment 1
Antibacterial peptide of the present invention (called after NKLP27) is the aminoacid sequence in sequence table SEQ ID No.1.
Sequence table SEQ ID No.1 is:
KVKARLIKICNKIGFLKSRCHKFVITH
(a) sequence signature:
● length: 27
● type: aminoacid sequence
● chain: strand
● topological framework: linearity
(b) molecule type: protein
(c) suppose: no
(d) antisense: no
(e) originate at first: Cynoglossus semilaevis
Constructional feature: be rich in basic aminoacids
Embodiment 2
The application of antibacterial peptide
Synthesizing of step 1) antibacterial peptide
The antibacterial peptide NKLP27 of above-described embodiment 1 is synthetic by " Shanghai Sangon Biological Engineering Technology And Service Co., Ltd ", purity >90%.
Step 2) antibacterial peptide injection
By above-mentioned steps 1) synthetic antibacterial peptide in PBS, be diluted to 20ug/ml, be antibacterial peptide diluent.20 Cynoglossus semilaevis (heavily about 10.1g) are divided into 4 groups, 5 every group at random.By these 4 groups difference called after A, B, C and D.Every fish of A and C group is injected respectively to 50ul antibacterial peptide diluent, every fish of B and D group (control group) is injected respectively to 50ul PBS.
Described PBS moiety is by weight percentage: 0.8%NaCl, 0.02%KCl, 0.358%Na
2hPO
4.12H
2o, 0.024%NaH
2pO
4, surplus is water.
Step 3) bacterium and viral suspension preparation
Culturing eel vibrio C312 to OD in LB substratum
600be 0.8, then centrifugal (5000g, 4 ℃, 10min), collects thalline, be suspended in PBS to final concentration be 10
6cfu/ml, is Vibrio anguillarum suspension.Concrete Megalocytivirus RBIV-C1(preparation method is shown in to Zhang M; Xiao Z; Hu Y; Sun L.Characterization of a megalocytivirus from cultured rock bream; Oplegnathus fasciatus (Temminck & Schlege), in China.Aquac Res.2012; 43:556 – 64) in PBS, be diluted to 10
5copies/ml, is viral suspension.
Described Vibrio anguillarum C312 is stored in the common micro-organisms center C GMCC of China Committee for Culture Collection of Microorganisms, depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, deposit number is: CGMCC No.6250, preservation date 2012.6.21, Classification And Nomenclature is Vibrio anguillarum (Vibrio anguillarum).
Step 4) is attacked poison and is infected
In above-mentioned steps 2) 1h after injection, by every fish injection 100ul above-mentioned steps 3 of A and B group) Vibrio anguillarum suspension, by every fish injection 100ul above-mentioned steps 3 of C and D group) viral suspension.Infect latter the 1st day on bacterium, get A and B group fish kidney and spleen tissue, homogenate in 2mlPBS, coats LB flat board by 100ul homogenate.Flat board is placed in to 30 ℃ and cultivates 48h, calculate the colony number occurring.After virus infection the 5th day, utilize DNA extraction test kit (be purchased from TIANGEN Biotech (Beijing) Co., Ltd. ") to extract DNA from C and D group fish kidney and spleen tissue, with absolute quantitation PCR method detect organize in viral level (concrete grammar is shown in above-mentioned reference).Result shows that the bacterial count of A group fish kidney and spleen (is respectively 2.1x10
2and 7.3x10
2) significantly (P<0.01) (be respectively 4.2x10 lower than the bacterial count of B group fish kidney and spleen
2and 2.5x10
3); The viral number of C group fish kidney and spleen (is respectively 2.5x10
3and 8.7x10
5) significantly (P<0.01) (be respectively 3.1x10 lower than the viral number of D group fish kidney and spleen
5and 4x10
6).
These results show, antibacterial peptide NKLP27 of the present invention can significantly strengthen infecting of fish opposing bacterium and virus.
Claims (2)
1. an antibacterial peptide, is characterized in that: antibacterial peptide is shown in the aminoacid sequence in sequence table SEQ ID No.1.
2. an application for antibacterial peptide described in claim 1, is characterized in that: described antibacterial peptide is for the preparation of the inhibitor of antibacterium or virus infection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410067697.4A CN103804482B (en) | 2014-02-26 | 2014-02-26 | A kind of antibacterial peptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410067697.4A CN103804482B (en) | 2014-02-26 | 2014-02-26 | A kind of antibacterial peptide and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103804482A true CN103804482A (en) | 2014-05-21 |
| CN103804482B CN103804482B (en) | 2015-09-23 |
Family
ID=50701911
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410067697.4A Expired - Fee Related CN103804482B (en) | 2014-02-26 | 2014-02-26 | A kind of antibacterial peptide and application thereof |
Country Status (1)
| Country | Link |
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| CN (1) | CN103804482B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109096383A (en) * | 2018-07-20 | 2018-12-28 | 青岛农业大学 | A kind of source of fish antibacterial peptide parasin I mutant and its application |
| CN111471094A (en) * | 2020-04-30 | 2020-07-31 | 大连理工大学 | Antimicrobial peptide MSPiscidin-2, and coding gene and application thereof |
| CN112970634A (en) * | 2021-02-04 | 2021-06-18 | 青岛农业大学 | Clown fish-vegetable symbiotic circulating water breeding method |
| CN113880933A (en) * | 2021-11-12 | 2022-01-04 | 青岛农业大学 | Antibacterial peptide SssNKL 27 and application thereof |
| CN114933643A (en) * | 2022-04-29 | 2022-08-23 | 华南农业大学 | Silurus meridionalis NKL (Natural killer L) antibacterial peptide as well as coding gene and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145822A (en) * | 2013-02-04 | 2013-06-12 | 中国科学院海洋研究所 | Fish natural killer (NK)-lysin effective factors and application method thereof |
-
2014
- 2014-02-26 CN CN201410067697.4A patent/CN103804482B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145822A (en) * | 2013-02-04 | 2013-06-12 | 中国科学院海洋研究所 | Fish natural killer (NK)-lysin effective factors and application method thereof |
Non-Patent Citations (3)
| Title |
|---|
| IKUO HIRONO ET.AL: "Characteriazation of Japanese flounder(Paralichthys olivaceus)NK-lysin,an antimicrobial peptide", 《FISH & SELLFISH IMMUNOLOGY》, vol. 22, no. 5, 30 May 2007 (2007-05-30), pages 567 - 575, XP005861817, DOI: doi:10.1016/j.fsi.2006.08.003 * |
| MIN ZHANG ET.AL: "A NK-lysin from Cynoglossus semilaevis enhances antimicrobial defense against bacterial and viral pahtogens", 《DEVELOPMENTAL & COMPARATIVE IMMUNOLOGY》, vol. 40, no. 34, 31 August 2013 (2013-08-31), pages 258 - 265 * |
| MIN ZHANG ET.AL: "NKLP27:A teleost NK-Lysin peptide that modulates immune response,induces degradation of bacterial DNA,and inhibits Bacterial and viral infection", 《PLOS ONE》, vol. 9, no. 9, 2 September 2014 (2014-09-02), pages 1 - 9 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109096383A (en) * | 2018-07-20 | 2018-12-28 | 青岛农业大学 | A kind of source of fish antibacterial peptide parasin I mutant and its application |
| CN109096383B (en) * | 2018-07-20 | 2021-05-04 | 青岛农业大学 | Fish-derived antibacterial peptide parasin I mutant and application thereof |
| CN111471094A (en) * | 2020-04-30 | 2020-07-31 | 大连理工大学 | Antimicrobial peptide MSPiscidin-2, and coding gene and application thereof |
| CN111471094B (en) * | 2020-04-30 | 2022-04-12 | 大连理工大学 | Antimicrobial peptide MSPiscidin-2, and coding gene and application thereof |
| CN112970634A (en) * | 2021-02-04 | 2021-06-18 | 青岛农业大学 | Clown fish-vegetable symbiotic circulating water breeding method |
| CN113880933A (en) * | 2021-11-12 | 2022-01-04 | 青岛农业大学 | Antibacterial peptide SssNKL 27 and application thereof |
| CN113880933B (en) * | 2021-11-12 | 2023-10-20 | 青岛农业大学 | Antibacterial peptide SsNKL27 and application thereof |
| CN114933643A (en) * | 2022-04-29 | 2022-08-23 | 华南农业大学 | Silurus meridionalis NKL (Natural killer L) antibacterial peptide as well as coding gene and application thereof |
| CN114933643B (en) * | 2022-04-29 | 2023-09-08 | 华南农业大学 | Silurus meridionalis NKL antibacterial peptide, and coding gene and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103804482B (en) | 2015-09-23 |
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