A recombinant s1 protein and its subunit vaccine of a new mutant strain of porcine epidemic diarrhea virus

A technology for porcine epidemic diarrhea and virus mutation, which can be used in antiviral agents, recombinant DNA technology, and cells modified by introducing foreign genetic material, which can solve the problems of not achieving good immune effects
CN104046637BActive Publication Date: 2017-04-12NANJING AGRICULTURAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
NANJING AGRICULTURAL UNIVERSITY
Publication Date
2017-04-12

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Abstract

The invention discloses a recombinant S1 protein of a novel mutant strain of a porcine epidemic diarrhea virus and a subunit vaccine of the recombinant S1 protein. The protein is obtained by the steps of cloning a fusion gene fragment PE(delta III)-S1(m)-KDEL3 containing a KDEL3 gene sequence, a gene sequence of III-region-deleted pseudomonas aeruginosa exotoxin A and an S1(m) gene sequence to a baculovirus vector to obtain a recombinant vector; carrying out recombinant vector site specificity transposition on a seroconversion DH10Bac competent cell of the recombinant vector to obtain a recombinant bacmid; transfecting the recombinant bacmid to an Sf9 cell under the mediation of a liposome to obtain a recombinant Sf9 cell; expressing by using the recombinant Sf9 cell. According to the invention, a PE(delta III)-S1(m)-KDEL3 fusion protein is firstly expressed by using a baculovirus expression system on the basis of optimizing the predilection of a codon of mammalian with S1 genes, so that the recombinant baculovirus for efficiently and accurately expressing the fusion protein is obtained. Proved by IFA and Western blot, the recombinant baculovirus can be used for accurately expressing the fusion protein, and the fusion protein has favorable reactionogenicity.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a recombinant S1 protein of a new mutant strain of porcine epidemic diarrhea virus and a subunit vaccine thereof. Background technique

[0002] Porcine epidemic diarrhea virus (PEDV) is a single-stranded positive-sense RNA virus with an envelope, belonging to type I coronavirus. Like other coronaviruses, the PEDV S protein plays an important role in inducing neutralizing antibodies, binding cells, and promoting virus and cell fusion [2] . Studies have shown that most of the neutralizing epitope regions of the S protein are located in the S1 region; in addition, comparing the PEDV mutant strains and traditional strains, the S protein has the largest variation, and the main variation occurs in the S1 region.

[0003] Pseudomonas exotoxin A (PEA) is a cytotoxic exotoxin secreted by Pseudomonas aeruginosa. PEA is composed of 613 amino acids and can be divided into three di...

Claims

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