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Lake Taihu whitefish host defense peptide and application thereof

A technology of host defense peptide and Taihu whitefish, which is applied in the field of biomedicine, can solve the problems of inferior immune system, perfection of higher animals, lagging sexual immune response, etc., and achieve the effect of low cost, small molecular weight and easy acquisition

Active Publication Date: 2018-08-31
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Teleosts also have a complete humoral and cellular immune response, but unlike mammals, the innate immune system of fish plays a more important role in the immune response against pathogen infection, mainly because fish are ectothermic animals, and the lymphocytes Proliferation is relatively slow, and the occurrence of acquired immune response is relatively delayed (up to 12 weeks). Secondly, the immune system of fish is not as perfect as that of higher animals in evolution, and the affinity of antibodies and the size of antibody library are significantly lower than those of mammals.

Method used

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  • Lake Taihu whitefish host defense peptide and application thereof
  • Lake Taihu whitefish host defense peptide and application thereof
  • Lake Taihu whitefish host defense peptide and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Sephadex Chromatography of Host Defense Peptides in Taihu Lake Whitefish Head Kidney

[0028] Taihu Lake whitefish was purchased from Taihu Lake Aquaculture Center, with a body length of 15-20 cm. The Taihu Lake whitefish was dissected, and the head kidney of the immune organ was collected, and 20mM PBS (1mL / 100mg tissue) containing 1% (v / v) protease inhibitor was added. Homogenize on ice, add liquid nitrogen, grind repeatedly three times, centrifuge at 5000g for 30min, collect supernatant, freeze and concentrate for later use. Filtration chromatography was performed using Sephadex G-50 Sephadex. Dissolve about 1g of lyophilized powder in 10mL of 0.1M Na 2 HPO 4 -NaH 2 PO 4 In the phosphate buffer solution of pH 6.0, load the sample on Sephadex G-50 (Superfine, GE healthcare) dextran gel filtration column (100cm×2.6cm) that has been equilibrated, elute with the same buffer solution, and use Collected by an automatic partial collector with a flow rate of ...

Embodiment 2

[0030] Example 2: Anti-aquatic activity of each elution peak of Sephadex G-50 dextran gel filtration chromatography detected by inhibition zone method

[0031] Firstly, the antibacterial zone method was used to detect the antibacterial activity of five elution peaks of the immune organ head kidney of Taihu Lake whitefish after Sephadex G-50 dextran gel filtration chromatography on Vibrio parahaemolyticus and Vibrio harveyi active. These two strains of aquaculture pathogenic bacteria were inoculated on the solid medium of nutrient broth, and cultured upside down in a 37°C incubator. After the colony grows, use an inoculation loop to pick a single colony and spread it evenly on the surface of the nutrient broth solid medium, paste a sterilized circular filter paper piece with a diameter of 0.5 cm on the surface of the medium, and take the sample solution to be tested (10μL, about 2mg / mL) was added dropwise on the filter paper. Incubate upside down in a 37°C incubator for 18-20...

Embodiment 3

[0033] Embodiment 3: reversed phase high performance liquid chromatography

[0034]First, the third elution peak in Sephadex chromatography was further separated and purified by reverse phase-high performance liquid chromatography (RP-HPLC, C18, 25×0.46cm). An elution system composed of water (containing 0.1% trifluoroacetic acid): acetonitrile (containing 0.1% trifluoroacetic acid) was used for gradient elution at 0.7mL / min, and the light absorption at 220nm was monitored by ultraviolet light, and the peaks were collected and vacuum-frozen Concentration, inhibition zone method to track and detect antibacterial activity.

[0035] The result is as image 3 As shown, the third elution peak after dextran gel filtration chromatography was divided into about 13 elution peaks after reversed-phase high performance liquid chromatography, and each elution peak was collected and vacuum freeze-dried. According to the filter paper method in the previous step, the activity of anti-aquacu...

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Abstract

The invention discloses a Lake Taihu whitefish host defense peptide and application thereof and belongs to technical field of biomedicines. The Lake Taihu whitefish host defense peptide disclosed by the invention is easy to obtain, in addition low in cost and small in molecular weight and can be prepared through chemical synthesis; an encoding gene of the peptide is cloned, and the peptide can beprepared by establishing a pronucleus or eukaryotic expression vector through large-scale fermentation; in addition, the Lake Taihu whitefish host defense peptide has a killing function on aquaculturepathogenic bacteria, particularly drug-resistant aquaculture pathogenic bacteria, in addition is a constitution part of an immune system of Lake Taihu whitefish self, and can be prepared into a novelimmune preparation to be applied to aquaculture of Lake Taihu whitefish.

Description

technical field [0001] The invention relates to a Taihu whitefish host defense peptide and its application, belonging to the technical field of biomedicine. Background technique [0002] The scientific name of the white fish in Taihu Lake is Erythroculter ilshaeformis (topmouth culter: Erythroculter ilshaeformis), and it has many common names: great white fish, cocked mouth, cocked shell, white silk, Xingkai big white fish, cocked culter, stinging fish, cocked white, white Fish, pouting silver carp, etc., are commonly known as Yangtze River Heshun in Guangdong, white fish in the middle reaches of the Yangtze River, and white fish in Taihu Lake in the lower reaches of the Yangtze River. It belongs to the family Cyprinidae, the subfamily of Cyprinidae, and the genus Cyprinidae. It is the largest and fastest-growing species of fish in the genus Cyprini. Taihu Lake white fish is white and tender, with delicious taste. It is one of the high-quality and important economic fish in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C12N15/12A61K38/17A61P31/04A23K20/147A23K50/80
CPCA23K20/147A23K50/80A61K38/00A61P31/04C07K14/461
Inventor 卫林徐薇冯婷婷刘天翌
Owner SUZHOU UNIV
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