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Enterocytozoon hepatopenaei spore wall protein as well as preparation and application of polyclonal antibody thereof

A polyclonal antibody, spore wall protein technology, applied in the direction of antibodies, immunoglobulins, serum immunoglobulins, etc., can solve the problems of harm, difficult to detect in time, timely treatment and effective control.

Inactive Publication Date: 2020-11-10
CHONGQING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

That is to say, after shrimps are infected with E. hepatica, there are no obvious symptoms in the early stage, and it is difficult to detect, treat and effectively control them in time, which has brought great harm to the shrimp farming industry in my country and other Asian countries.

Method used

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  • Enterocytozoon hepatopenaei spore wall protein as well as preparation and application of polyclonal antibody thereof
  • Enterocytozoon hepatopenaei spore wall protein as well as preparation and application of polyclonal antibody thereof
  • Enterocytozoon hepatopenaei spore wall protein as well as preparation and application of polyclonal antibody thereof

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Experimental program
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Embodiment

[0026] 1. Construction of protein EHP00_430 prokaryotic expression plasmid

[0027] Genomic DNA of Shrimp Enteroplasma hepatica was extracted as a template, EHP00_430 Gene-specific primers (430-F / 430-R) were used for PCR amplification to obtain the target band with a size of 750 bp. The target band was excised under ultraviolet light for gel recovery, and the recovered target gene fragment was first transformed into pMD19- T vector, and then through BamHI, SalI double digestion, the EHP00_430 The gene fragment was transferred into the pET28a vector to construct the recombinant prokaryotic expression plasmid, which was verified by sequencing.

[0028] 2. Prokaryotic expression and purification of protein EHP00_430

[0029] 2.1 Prokaryotic expression

[0030] Transform the recombinant plasmid and pET28a empty vector into Escherichia coli Rosetta (DE3) competent cells, spread on a plate containing corresponding antibiotics after heat shock; randomly pick 3-5 single colonies ...

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Abstract

The invention provides an enterocytozoon hepatopenaei spore wall protein as well as preparation and application of a polyclonal antibody thereof, and belongs to the technical field of animal quarantine. The enterocytozoon hepatopenaei spore wall protein (EHP00_430) is positioned on the surfaces of enterocytozoon hepatopenaei spores in the infection period, has high abundance, plays an important role in the process that enterocytozoon hepatopenaei infects prawns, and is an important target for detection, prevention and control of enterocytozoon hepatopenaei. The enterocytozoon hepatopenaei spore wall protein provided by the invention is used for preparing a polyclonal antibody for resisting the EHP00_430 protein; and the polyclonal antibody is high in specificity, can be used for accuratelydetecting enterocytozoon hepatopenaei in the infection period, and can be applied to detection of enterocytozoon hepatopenaei pathogens.

Description

technical field [0001] The invention relates to the technical field of animal quarantine, in particular to the preparation and application of a shrimp hepatic Enterospora sporozoite protein and a polyclonal antibody thereof. Background technique [0002] Microsporidia (Microsporidia) is a single-celled eukaryotic organism specializing in intracellular parasitism. It has been found that 150 genera and more than 1,400 species have a very wide range of hosts, including humans and many economic animals such as shrimps, crabs, rodents, etc. main pathogens. Shrimp Enterococcus hepatica ( Enterocytozoon hepatopenaei , abbreviated as EHP) is a new microsporidian pathogen belonging to the family Microsporidiaceae and the genus Enterococcus, which can infect Litopenaeus vannamei. At present, it has been found in Thailand, Indonesia, India, Vietnam, Malaysia, China and other places that the infection of shrimp Enterocystoma hepatica leads to slow or stagnant growth of cultured specie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/44C07K16/20C07K16/06G01N33/569A61K39/395A61P33/02
CPCA61P33/02C07K14/44C07K16/06C07K16/065C07K16/20C07K2317/76G01N33/56905G01N2469/10
Inventor 范晓东周泽扬杨小娟魏春梅肖嫒
Owner CHONGQING NORMAL UNIVERSITY
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