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SWP2 protein-resistant monoclonal cell strain and application thereof

A monoclonal and cell line technology, applied in the direction of immunoglobulin, equipment, peptides, etc., can solve the problems of slow growth, no growth, empty feed, etc., and achieve the effect of high affinity

Active Publication Date: 2020-05-12
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the disease was difficult to detect earlier, it often takes more than 30-45 days to show slow growth and no growth, so it is very easy to cause empty feed consumption and bring greater economic losses to farmers

Method used

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  • SWP2 protein-resistant monoclonal cell strain and application thereof
  • SWP2 protein-resistant monoclonal cell strain and application thereof
  • SWP2 protein-resistant monoclonal cell strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0024] 1 Construction of SWP2 recombinant expression plasmid

[0025] According to the genome information of SWP2, the PCR product of the SWP2 gene fragment was obtained by gene synthesis method; the PCR product and the expression vector Pet15B were double-digested with NdeI and XhoI, the double-digested fragment and the carrier were recovered by tapping the glue, and the two were connected with T4DNAigase; The connection solution was transferred into top10 competent cells, positive clones were screened and identified by sequencing.

[0026] 2.1 Expression and purification of SWP2 recombinant protein

[0027] 2.1 Expression

[0028] Transform the recombinant plasmid into Escherichia coli competent cells, spread it on a plate containing the corresponding antibiotic after heat shock, and cultivate it; pick a single clone and culture it in a liquid medium containing antibiotics, and add an inducer when the OD value reaches 0.6 IPTG continued to cultivate, and cultured at 20°C f...

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PUM

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Abstract

The invention discloses an SWP2 protein-resistant monoclonal cell strain and an application thereof, and belongs to the technical field of agricultural and animal quarantine. On one hand, the invention provides a novel SWP2 protein-resistant monoclonal cell strain and a SWP2 monoclonal antibody secreted by the monoclonal cell strain, and on the other hand, the invention provides the application ofthe monoclonal cell strain and the SWP2 monoclonal antibody in preparing a reagent for detecting enterocytozoon hepatopenaei diseases. The monoclonal antibody generated by the SWP2 protein-resistantmonoclonal cell strain provided by the invention can be combined with an SWP2 antigen with high specificity, has high affinity, and can provide accurate results for judging whether shrimp bodies are infected with enterocytozoon hepatopenaei (EHP) or not.

Description

technical field [0001] The invention belongs to the technical field of agriculture and animal quarantine, and specifically relates to a monoclonal cell strain against SWP2 protein and its application, and the SWP2 monoclonal antibody produced by the monoclonal cell strain and its application. Background technique [0002] Microsporidia (microsporidia) are single-celled eukaryotic endoparasites that widely parasitize shrimp, crustaceans and other animals. Microsporidia hepatica of prawns ( Enterocytozoon hepatopenaei , EHP) is a kind of microsporidia, which belongs to Microsporidiaceae and Enterocystis genus. It was first cultured in Thailand in 2009. Monodon ( Penaeus monodon ), and then EHP was detected successively in the culture areas of Litopenaeus vannamei, White prawn and Penaeus prawn in various Asian countries. Studies have shown that EHP has a wide range of infection routes. It can be transmitted horizontally through polluted aquaculture water and diseased shrimp...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/20G01N33/577G01N33/569
CPCC07K16/20G01N33/577G01N33/56905
Inventor 沈卫锋牛宝龙楼宝
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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