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Vaccine composition and its preparation method and application

A vaccine composition and the technology of the composition are applied in the directions of botanical equipment and method, biochemical equipment and method, application, etc., which can solve the problems of weak immunogenicity, low production efficiency and high production cost, and achieve good immune effect. , the effect of short time-consuming and simple preparation method

Active Publication Date: 2021-07-23
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Subunit vaccines do not contain nucleic acid substances, are relatively safe, and will not produce persistent or latent infection after vaccination. The immune response produced can be distinguished from wild virus infection, which is conducive to the control and elimination of the disease. However, the current research on bovine infection The production cost of rhinotracheitis virus subunit vaccine is high, the production efficiency is low, and the immunogenicity is not as good as attenuated vaccine and inactivated vaccine, so the application is limited

Method used

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  • Vaccine composition and its preparation method and application
  • Vaccine composition and its preparation method and application
  • Vaccine composition and its preparation method and application

Examples

Experimental program
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Effect test

Embodiment approach

[0027] As an embodiment of the present invention, the nucleotide sequence encodes a protein with the amino acid sequence shown in SEQ ID NO.2 in the Sequence Listing.

[0028] The second aspect of the present invention is to provide a bovine infectious rhinotracheitis virus gB protein fragment encoded by the nucleotide sequence of claim 1, or a conservative variant thereof, or an active fragment thereof.

[0029] As an embodiment of the present invention, the gB protein fragment is the amino acid sequence shown in SEQ ID NO: 2 in the sequence table.

[0030] The term "gB protein", also known as "glycoprotein B", is the most conserved glycoprotein among herpesvirus members.

[0031] The term "gD protein", also known as "glycoprotein D", is an essential structural protein for bovine infectious rhinotracheitis virus infection, and is one of the main glycoproteins on the membrane surface of mature virions.

[0032] As an embodiment of the present invention, the gB protein fragmen...

Embodiment 1

[0080] Example 1 Tandem expression of bovine infectious rhinotracheitis virus truncated gB protein and gD protein

[0081] 1. Synthesis of gB protein gene fragment and gD protein gene of bovine infectious rhinotracheitis virus

[0082] According to IBR Cooper's sequence (accession number: KU198480.1) the gB protein and gD protein gene sequences were designed and synthesized, and the gene synthesis was carried out by a gene synthesis company, and baculovirus was added to the N-terminal of the gB protein and gD protein gene sequences. The signal peptide of GP67 protein, with 6 histidine tags added to the C-terminus. The synthesized gB protein gene fragment is shown in sequence SEQ ID NO: 1, and the gD protein gene is shown in sequence SEQ ID NO: 3.

[0083] 2. Construction of tandem expression donor plasmids for truncated gB protein and gD protein

[0084] The gD protein gene synthesized in step 1 was double-digested with KpnI+NcoI, ligated with pFastBac Dual that had undergon...

Embodiment 2

[0093] The preparation of embodiment 2 bovine infectious rhinotracheitis virus truncated gB protein

[0094] 1. Construction of Donor Plasmids

[0095] Using the gB protein gene fragment synthesized in Example 1, the synthesized gB protein gene fragment was double digested with BamHI and EcoRI, connected to the pFastBacI vector that had undergone the same double digestion, and transformed into DH5α competent cells, and the correct positive plasmid was identified Named pFastBac-co-gBdel.

[0096] 2. Construction of recombinant bacmid

[0097] Referring to the construction method of the recombinant bacmid in Example 1, the donor plasmid obtained in step 1 of this example was used to construct the recombinant bacmid, and the correct recombinant bacmid identified by PCR was named Bacmid-co-gBdel.

[0098] 3. Acquisition and passage of recombinant baculovirus

[0099] With reference to the method for obtaining and subculturing the recombinant baculovirus in Example 1, the correct ...

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Abstract

The present invention provides a gB protein fragment of bovine infectious rhinotracheitis virus, or a conservative variant thereof, or an active fragment thereof. The gB protein fragment has a high expression level, and the prepared subunit antigen immunization effect is better than that of the gB protein. The invention also provides a subunit vaccine prepared by tandemly expressing the gB protein fragment and the gD protein. The vaccine preparation method is simple and has better protective effect against bovine infectious rhinotracheitis virus caused by bovine infectious rhinotracheitis virus.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and specifically relates to a subunit vaccine composition, a preparation method thereof, and an application of the vaccine composition. Background technique [0002] Infectious Bovine Rhinotracheitis Virus (IBRV), also known as Bovine Herpesvirus 1 (BHV-1), is an important pathogen of cattle, which causes bovine infectious rhinotracheitis, It is an acute, febrile, contact infectious disease of cattle. The decreased cellular immunity and immunosuppression caused by bovine infectious rhinotracheitis virus often lead to secondary bacterial infections, making the clinical diagnosis of the disease difficult. The World Organization for Animal Health (OIE) lists bovine infectious rhinotracheitis as a category B animal disease. [0003] Vaccination is one of the main measures to prevent, control and even eliminate bovine infectious rhinotracheitis. Subunit vaccines do not contain nucleic ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/38C07K14/06C07K19/00A61K39/265A61P31/22
CPCA61K39/12C07K14/005C07K2319/00C12N2710/16022C12N2710/16034
Inventor 田克恭王同燕孙进忠张许科
Owner PU LIKE BIO ENG
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