PCV3 epitope polypeptide sequence screening method

A technology of antigenic epitopes and polypeptide sequences, applied in the field of immunology, can solve the problems of no diagnosis, prevention and treatment measures, and achieve good antigenicity

Inactive Publication Date: 2018-07-13
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
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Problems solved by technology

Due to the outbreak of PCV3 epidemic, there is no relevant diagnosis, prevention and treatment measures

Method used

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  • PCV3 epitope polypeptide sequence screening method
  • PCV3 epitope polypeptide sequence screening method
  • PCV3 epitope polypeptide sequence screening method

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Embodiment

[0035] The screening of the polypeptide sequence of PCV3 epitope comprises the following steps:

[0036] First, according to the amino acid sequence of the known PCV3 cap protein published by NCIBI, the amino acid sequence was compared and analyzed to observe the conservation of its amino acid;

[0037] Secondly, DNASTAR7.0 software was used to predict the amino acid antigenicity of the cap protein of one of the PCV3 strains, and the prediction results are shown in figure 1 shown;

[0038] Then, by PCR method, the predicted five short peptide sequences were respectively amplified, and cloned into the prokaryotic expression vector pGEX-4T-3, and transformed into BL21 (DE3) to induce expression; the induced expression The bacteria were ultrasonically disrupted, using GST protein purification column, after the purified protein was subjected to SDS-PAGE electrophoresis, the electrophoresis results were as follows figure 2 As shown, 1-5 are 5 short peptide samples respectively, ...

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Abstract

The invention discloses a PCV3 epitope polypeptide sequence screening method. The amino acid sequences of comprised five polypeptides are respectively PCV3-1: GTPQNNKPWH; PCV3-2: SPAQQTKTMF; PCV3-3: WLQDDPYAESSTRKV; PCV3-4: MTSKKKHSRY; PCV3-5: VPEKTGMTDFYGTKE. The method uses an ELISA (Enzyme-Linked Immuno Sorbent Assay) method to detect antiserums IgG of PCV2 and PCV3, and an oligopeptide specifically combined with a PCV3 antibody is screened through the ELISA method. The screened oligopeptide provides a reliable evidence for a PCV3 subunit vaccine, a related agent used for diagnosing the infection of a circovirus 3, and an antibody for treating the infection of the circovirus 3, etc. The PCV3 epitope polypeptide sequence screening method has the advantages that the antigenicity of the screened five polypeptides are good, the epitope is identified to be held by the PCV3, the sequence is conservative, whether an animal is infected with the PCV3 is identified effectively by applying theepitope sequence, and an effective detection means is provided for the prevention of the PCV3 in future.

Description

technical field [0001] The invention belongs to the technical field of immunology, and specifically relates to a polypeptide sequence screening method capable of identifying PCV3 epitopes of antibodies after PCV2 and PCV3 infection. Background technique [0002] Circovirus is very harmful to my country's pig industry, and it can be mixed with various pathogens, which can easily cause immune failure. At present, PCV2 infection is more clinically harmful, but in recent years, circovirus has recombined and mutated in the body, and a new type, PCV3, has emerged. Relevant data show that PCV3 infection alone can cause morbidity and even death in pig herds, and mixed infection with PCV2 can cause greater damage to pig herds. [0003] However, since PCV3 is a recently discovered virus strain, there is no clinically effective prevention and control method. During the evolution process of the virus, in order to escape the immune surveillance of the body, it is bound to mutate releva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/01G01N33/68G01N33/569
CPCC07K14/005C12N2750/10022C12N2750/10034G01N33/56983G01N33/68
Inventor 蒋再学罗满林刘向楠吴梦凡霍礼侠吴佳俊
Owner SOUTH CHINA AGRI UNIV
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