Porcine circovirus type 3 genetic engineering subunit vaccine and preparation method thereof

A subunit vaccine, porcine circovirus technology, applied in the fields of biopharmaceuticals and genetic engineering, can solve the problems of low protection possibility and no protective effect of the virus

Active Publication Date: 2018-11-16
荣俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Given the low likelihood of cross-protection between PCV2 and PCV3, currently available vaccines are not protective against this virus

Method used

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  • Porcine circovirus type 3 genetic engineering subunit vaccine and preparation method thereof
  • Porcine circovirus type 3 genetic engineering subunit vaccine and preparation method thereof
  • Porcine circovirus type 3 genetic engineering subunit vaccine and preparation method thereof

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Embodiment Construction

[0045] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and through specific implementation methods.

[0046] For better illustrating the present invention, facilitate understanding technical scheme of the present invention, typical but non-restrictive embodiment of the present invention is as follows:

[0047] 1. Construction of PCV3 cap protein gene cloning vector

[0048] 1. Materials and methods

[0049] 1.1. Extraction of disease material DNA

[0050] Get 0.5 g of the inguinal lymph node sample of dead pigs after porcine circovirus type 3 infection, add 4.5 mL of sterile normal saline, get 472 μ L of lymph node homogenate after fully homogenizing, add 25 μ L of 10% SDS (sodium dodecyl sulfate) and 2.5 μL of proteinase K (25 mg / ml), placed in a 50°C water bath for 2 hours, added 500 μL of Tris-saturated phenol, vortexed for 30 seconds, and centrifuged at 12,000 rpm for 10 minutes. Transfer the...

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Abstract

The invention provides a porcine circovirus type 3 (PCV3) genetic engineering subunit vaccine and a preparation method thereof. The method comprises the following steps: S1, constructing a PCV3cap protein gene cloning vector; S2, carrying out splicing and expression screening on a PCV3cap protein gene cloned by S1, and determining a splicing manner; S3, optimizing a codon for expressing the PCV3cap protein gene screened by S2; S4, fusing and connecting a specific gene segment with the upstream of a PCV3cap protein gene N-end gene optimized by S3; S5, constructing expression engineering bacteria by utilizing the gene recombined by S4, and expressing soluble PCV3cap protein by adopting the expression engineering bacteria, wherein the subunit molecular weight is about 25 kDa; purifying the soluble PCV3cap protein; S6, obtaining purified protein which is circovirus sample particles with the diameter of 17.74 nm to 18.20 nm under an electron microscope, and adding an immunoenhancer into thepurified expression protein to prepare the vaccine. After experiment pigs are immunized by utilizing the vaccine, an antibody, which resists the PCV3cap protein, can be detected, and the PCV3 virus loading amount in blood serum of the experiment pigs can be remarkably reduced in a anti-virus protection experiment; the vaccine has a remarkable protection effect on the experiment pigs.

Description

technical field [0001] The invention relates to the fields of genetic engineering and biopharmaceuticals, and relates to a porcine circovirus type 3 genetic engineering subunit vaccine and a preparation method thereof, in particular to a recombinant porcine circovirus type 3 genetic engineering subunit vaccine and a preparation method thereof. Background technique [0002] Porcine Circovirus (Porcine Circovirus, PCV) belongs to Circoviridae Circovirus genus, the virus has no envelope, is a single-strand negative-sense circular DNA virus, and is one of the smallest animal viruses. The traditional point of view is that there are two genotypes of porcine circovirus: porcine circovirus type 1 (PCV1) and porcine circovirus type 2 (PCV2), wherein PCV2 has pathogenicity. In June 2015, a porcine dermatitis nephropathy syndrome (PDNS) outbreak broke out in a pig farm in North Carolina, USA. Compared with the historical average, the mortality rate of sows increased by 10.2%, and the p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N15/34C12N1/21C07K14/01C07K19/00A61K39/12A61P31/20C12R1/19
CPCA61K39/12A61P31/20C12N15/70C07K14/005A61K2039/5258A61K2039/552A61K2039/55505C07K2319/00C12N2800/22C12N2750/00022C12N2750/10034
Inventor 荣俊匡红艳李国攀
Owner 荣俊
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