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Preparation method of flavobacterium columnare genetic recombination vaccines

A Flavobacterium columnar and gene recombination technology, applied in the field of molecular biology and aquatic products, can solve problems such as poor stability, potential safety hazards, and loss of protein immunogenicity, so as to reduce fishery economic losses, stabilize immune effects, and produce high efficiency effect

Active Publication Date: 2014-01-08
INST OF AQUATIC LIFE ACAD SINICA +1
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0009] Flavobacterium columnar inactivated vaccine has complex components, low stability, and the inactivation process will lead to the loss of immunogenicity of some proteins and affect its immune effect
Although the live attenuated Flavobacterium columnar vaccine has a good effect, the live attenuated vaccine has the risk of reverting mutation and has potential safety hazards

Method used

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  • Preparation method of flavobacterium columnare genetic recombination vaccines
  • Preparation method of flavobacterium columnare genetic recombination vaccines
  • Preparation method of flavobacterium columnare genetic recombination vaccines

Examples

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Embodiment 1

[0034] 1. Construction of expression vector

[0035] (1) Download the gene sequences of five genes of Flavobacterium columnar zinc protease, proline oligopeptidase, thermophilic protease, collagenase, and chondroitin sulfate AC lyase from Genbank, and the transmembrane region, hydrophilic region and antigen The region was analyzed, and the primers shown in Table 1 were designed and synthesized.

[0036] Primers used in Table 1

[0037]

[0038] (2) The genome of Flavobacterium columnarum (the strain is commercially available or can be obtained from China General Microorganism Collection Center) was extracted using the bacterial genome extraction kit of OMEGA Company.

[0039] (3) The expression fragment was amplified by PCR using the genomic DNA of Flavobacterium columnar as a template.

[0040] (4) The size of the PCR amplification product was identified by 1.0% agarose gel electrophoresis (see figure 1 ), DNA fragments were recovered using an agarose gel recovery kit, ...

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Abstract

The invention discloses a preparation method of flavobacterium columnare genetic recombination vaccines. The preparation method of flavobacterium columnare genetic recombination vaccines comprises the steps of (1) constructing expression carriers; (2) expressing and purifying target protein; (3) adjusting the concentration of the purified protein so as to obtain the flavobacterium columnare genetic recombination subunit vaccines which can be used directly. The vaccines prepared by the invention have the advantages of high safety, stable immune effect, high production efficiency and the like.

Description

technical field [0001] The invention relates to the fields of molecular biology and aquatic products, in particular to a vaccine preparation method capable of preventing grass carp from being infected by Flavobacterium columnar. Background technique [0002] Flavobacterium columnar is widely distributed and is a worldwide pathogenic bacteria of aquatic animals. Its host range is extremely wide, and it can infect fish of many families including Cyprinidae, Perch, Catfish, Salmonidae, Sunidae, and Mulletidae. Almost all freshwater fish are sensitive to it. Under aquaculture or natural conditions Sea fish and ornamental fish can also be infected. In the United States, Flavobacterium columnar is the second most important pathogen affecting channel catfish farming after Edwardsiella, causing economic losses of about $1 million per year. In my country, Flavobacterium columnar mainly harms grass carp, mandarin carp, carp, crucian carp and other fishes, and the columnar disease ca...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P31/04C12N15/74
Inventor 聂品罗璋刘志新
Owner INST OF AQUATIC LIFE ACAD SINICA
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