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White spot syndrome virus gene VP28, recombinant protein, polyclonal antibody, preparation method and application

A white spot syndrome, polyclonal antibody technology, applied in the direction of antiviral immunoglobulin, virus/phage, botanical equipment and methods, etc., can solve problems such as inability to perform WSSV determination, lack of Procrayfish, etc.

Inactive Publication Date: 2020-12-18
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the above-mentioned technical problems, the present invention provides white spot syndrome virus VP28 gene, recombinant protein, polyclonal antibody and preparation method and application, to solve the problems such as the lack of procambarus clarkii WSSVVP28 detection antibody and the inability to measure WSSV

Method used

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  • White spot syndrome virus gene VP28, recombinant protein, polyclonal antibody, preparation method and application
  • White spot syndrome virus gene VP28, recombinant protein, polyclonal antibody, preparation method and application
  • White spot syndrome virus gene VP28, recombinant protein, polyclonal antibody, preparation method and application

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Embodiment 1

[0046] 1. WSSV Induced expression of VP28-His recombinant protein:

[0047] Competent cells E. coli Add pET-B2M- WSSV The VP28 recombinant plasmid was mixed gently, left to stand in an ice bath for 5 min, then placed in a 42°C water bath for 90 s heat shock, and quickly transferred to an ice bath. Add 900 μL of sterile LB medium to each centrifuge tube, mix well, place on a shaker at 37°C for 45 min, and resuscitate the bacteria, then draw 100 μL of the recovered bacteria solution and spread it on the culture medium containing kanapenicillin Resistant LB solid medium was cultured at 37°C for 12-16 h. Select the Escherichia coli containing the recombinant plasmid, add liquid medium, shake culture at 37°C until the OD600 of the bacterial liquid is 0.4-0.6, add an appropriate amount of IPTG to a final concentration of 1 mM, induce expression for 4 hours, collect the bacterial cells, and resuspend in sterilized water Bacterial cells were stored at -80°C and frozen and thawed...

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Abstract

The invention belongs to the field of genetic engineering of molecular biology, relates to a nucleotide sequence of a procambarus clarkii white spot syndrome virus gene VP28, an amino acid sequence encoded by the nucleotide sequence and a codon-optimized nucleotide sequence and particularly relates to a preparation method for recombinant expression of a protein WSSVVP28 and preparation of a rabbitpolyclonal antibody. Through the recombinant-expressed protein WSSVVP28, a structure and functions of the protein can be further analyzed in vitro; and furthermore, due to the preparation of the polyclonal antibody, a foundation can be settled for further detecting the content of WSSV in procambarus clarkii bodies.

Description

technical field [0001] The nucleotide sequence and coded amino acid sequence of the VP28 gene of WSSV, as well as the nucleotide sequence after codon optimization, especially relate to the recombinant expression preparation method of the VP28 protein of WSSV and the preparation of the anti-rabbit polyclonal antibody. The structure and function of the protein can be further analyzed in vitro by recombinantly expressing the VP28 protein of WSSV. In addition, the preparation of polyclonal antibodies can lay the foundation for further detection of the content of WSSV in fish and shrimp. Background technique [0002] VP28 of WSSV is the envelope protein of white spot syndrome virus that has been studied the most at present. The protein is one of the most important components of the virus, including structural proteins and non-structural proteins. The structural proteins of WSSV include three parts: envelope protein, nucleocapsid protein and envelope protein. They maintain the bas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/34C07K14/01C07K16/08G01N33/68G01N33/569C12N15/70C07K16/06
CPCC07K14/005C07K16/06C07K16/081C12N15/70C12N2710/14022C12N2800/22G01N33/56983G01N33/6854G01N2333/01G01N2469/10G01N2469/20
Inventor 杨辉熊浩然张莹莹魏文志
Owner YANGZHOU UNIV
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