Vitellus immune globulin for preventing prawn virus, tis preparing method and use thereof

A yolk immunoglobulin, virus disease technology, applied in the direction of antiviral immunoglobulin, egg-derived immunoglobulin, antiviral agent, etc. The effect is weakened, etc., to achieve the effect of improving disease resistance, safe use, and direct effect.

Inactive Publication Date: 2005-02-23
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 1. The pathogen of prawn white spot syndrome is a type of virus. There are differences in the white spot disease virus infected by different hosts in different regions. Only one kind of white spot syndrome virus is selected to prepare the antigen. It cannot be applied to infections caused by other virus strains, the actual application range is narrowed, and the effect of inhibiting and killing viruses is weakened
[0010] 2. The nucleic acid immunogen is made without using the white spot syndrome virus gene or its antigenic determinant gene, which brings

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0035] Example 1

[0036] Preparation of crawfish primary cells

[0037] Take healthy crayfish without WSBV infection, clean the body surface, soak in 0.01% potassium permanganate solution for 5 minutes, rinse with sterile water, and disinfect the junction between the head and breastplate and abdomen with alcohol cotton ball. Use a sterile syringe to draw 2~3mL of anticoagulant (0.14mol / L NaCl, 0.1mol / L glucose, 30mmol / L citric acid, 26mmol / L sodium citrate, 10mmol / L EDTA, pH4.6), and collect blood from the sinuses , Anticoagulant and hemolymph were mixed and centrifuged at 1500r / min for 10min, the supernatant was discarded, the precipitate was washed twice with L-15 medium, and the whole culture solution (15% calf serum, 100U / ml penicillin and 100μg / ml streptomycin L-15 culture medium), gently and evenly resuspend the lymphocytes, inoculate them into culture flasks or culture plates, and incubate overnight at 27°C for later use.

Example Embodiment

[0038] Example 2

[0039] Target gene cloning

[0040] The VP28 gene was amplified by PCR. The sequencing results showed that the gene contained 615 nucleotides. The sequence fragments of WSSV from different sources in GeneBamk had a homology of 100%.

Example Embodiment

[0041] Example 3

[0042] Protein induced expression

[0043] SDS-PAGE electrophoresis results showed that the strain with the recombinant plasmid pGEX-2T produced a specific protein band at the molecular weight of 24KDa, 26KDa, and 28KDa, and its expression accounted for more than 40% of the total bacterial protein mass. , And no such corresponding protein bands were produced in the control strains.

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PUM

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Abstract

This invention refers to cultivating aquatic animal cultivating and, especially, to vitelline immune globulin' s preparation, which is capable to prevent prawn virus disease. The feature is that: preparing a kind mono-antigen and composite antigen with one or several kinds of inactivate strain / attenuated strain / gene subunit immunogen / nucleic acid immunogen, which belong to prawn leucoderma syndrome virus. The egg avian are uaed as biological reactors, immunity obtaining resisting prawn leucoderma syndrome a specificity LgY, which is alone or together with peptidoglycan, probiotics or Chinese herbs medicine, are capable of being active component of feedstuff additives, medicinal dipping agent, or prawn pool dousing agent to prevent from prawn diseases. The advantages are IgY having effect through passive immunity mechanism to inhibit or kill the virus directly and increase prawn 's immunological competence. It has obvious effect in treating leucoderma syndrome for prawn, and safety, environment protecting, convenient etc.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the technical field of aquatic animal breeding, and in particular relates to a yolk immunoglobulin (IgY) for preventing and treating prawn virus disease and a preparation method thereof. The IgY can be used as a feed additive or a medicine bathing agent or the active ingredient of the splashing agent to control the white spot syndrome of prawns. Background technique [0002] my country's prawn farming started in the late 1970s. The continuous improvement and perfection of aquaculture production technology made it quickly become one of the pillar industries of the mariculture industry. The density of stocking, coupled with the continuous deterioration of the ecological environment for farming, has made shrimp diseases, especially viral diseases of shrimps, an important factor hindering the development of this industry. From 1993 to 1994, prawn virus disease raged across my country's north a...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P31/12C07K16/02C07K16/08
Inventor 徐永平
Owner DALIAN UNIV OF TECH
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