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Bovine herpes virus antigen composition and application thereof

A technology of bovine herpes virus and composition, which is applied in the direction of virus antigen components, applications, viruses, etc., which can solve the problems of short immunization period, high cost of culling, and poor immunization effect

Active Publication Date: 2020-10-23
苏州世诺生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the global control of IBR mainly adopts two methods: culling and vaccination, but culling is costly, and general vaccines also have problems such as poor immune effect and short immunization period

Method used

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  • Bovine herpes virus antigen composition and application thereof
  • Bovine herpes virus antigen composition and application thereof
  • Bovine herpes virus antigen composition and application thereof

Examples

Experimental program
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preparation example Construction

[0047] The preparation method of bovine herpes virus recombinant protein according to one embodiment of the present invention comprises the following steps: cultivating the above-mentioned host cells under suitable conditions, collecting culture fluid and / or host cell lysates, and then separating and purifying to obtain recombinant bovine herpes virus protein.

[0048] In a specific example, separation and purification methods include nickel column affinity chromatography, molecular sieve chromatography, etc., are not limited thereto, and can be selected according to needs.

[0049] The bovine infectious rhinotracheitis vaccine according to one embodiment of the present invention comprises the above bovine herpes virus antigen composition and a pharmaceutically acceptable adjuvant.

[0050] In a specific example, the adjuvant can be one or a combination of two or more of MONTANIDE ISA 206 VG, MONTANIDE ISA 201 VG, liquid paraffin, camphor oil, lectin, etc., preferably MONTANID...

Embodiment 1

[0052] Example 1 Construction of recombinant eukaryotic expression vector pCI-gB-gD-GS

[0053] 1. Amplification and purification of gB-gD gene expression cassette

[0054] The codon-optimized gB-gD gene expression cassette (SEQ IDNO:9) was synthesized in Nanjing GenScript Biotechnology Co., Ltd. and cloned into the pUC-57 vector to obtain the pUC-gB-gD plasmid vector. PCR amplification was performed using pUC-gB-gD as a template and gB-gD-F and gB-gD-R as primers. The amplification system is shown in Table 1. The reaction conditions were: pre-denaturation at 94°C for 5 minutes; denaturation at 95°C for 45 seconds, renaturation at 60°C for 45 seconds, extension at 72°C for 2 minutes, 30 cycles; extension at 72°C for 10 minutes, and storage at 4°C.

[0055] gB-gD-F: 5'-ATAGGTACCgccgccaccatggaaaccgataccctgctgctgtgggtgctgctg-3'

[0056] gB-gD-R: 5'-ATACTCGAGttaatgatgatgatgatgatggccttccgggccGCAcgggccgccgttg-3'

[0057] Table 1 gB-gD gene expression cassette amplification system...

Embodiment 2

[0074] Example 2 Construction of recombinant eukaryotic expression vector pCI-gH-gL-GS

[0075] 1. Amplification and purification of gH-gL gene expression cassette

[0076] The codon-optimized gH-gL gene expression cassette (SEQ ID NO: 10) was synthesized in Shanghai Sunny Biotechnology Co., Ltd. and cloned into the pUC-57 vector to obtain the pUC-gH-gL plasmid vector. Using pUC-gH-gL as template, gH-gL-F, gH-gL-R as primers for PCR amplification, the amplification system is shown in Table 5. The reaction conditions were: pre-denaturation at 94°C for 5 minutes; denaturation at 95°C for 45 seconds, renaturation at 60°C for 45 seconds, extension at 72°C for 2 minutes, 30 cycles; extension at 72°C for 10 minutes, and storage at 4°C.

[0077] gH-gL-F: 5'-ATAGGTACCGCCGCCACCatggaaaccgataccctgctgctgtgggtgctgctg-3'

[0078] gH-gL-R: 5'-ATACTCGAGttaatgatgatgatgatgatggcgataaatgccatcgc-3'

[0079] Table 5 gH-gL gene expression cassette amplification system

[0080]

[0081] Perfor...

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Abstract

The invention relates to a bovine herpes virus antigen composition and an application thereof. The bovine herpes virus antigen composition comprises at least two of bovine herpes virus recombinant gBprotein having an amino acid sequence as shown in SEQ ID NO: 1, bovine herpes virus recombinant gD protein having an amino acid sequence as shown in SEQ ID NO: 2, bovine herpes virus recombinant gH protein having an amino acid sequence as shown in SEQ ID NO: 3 and bovine herpes virus recombinant gL protein with an amino acid sequence as shown in SEQ ID NO: 4. Sequence information and space structures of gB protein, gD protein, gH protein and gL protein of bovine herpes virus are comprehensively analyzed, site-specific mutagenesis is performed on the four proteins respectively, respective immunogenicity of the proteins is improved, neutralizing antibody can be better stimulated, the immune effect is better, and the immune period is longer.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a bovine herpes virus antigen composition and application thereof. Background technique [0002] Infectious Bovine Rhinotracheitis (IBR) is a highly contagious disease caused by Infectious Bovine Rhinotracheitis Virus (IBRV). The clinical symptoms caused by the disease mainly include dyspnea, rhinitis, fever, loss of appetite, conjunctivitis, mastitis, miscarriage, balanoposthitis, vulvovaginitis and systemic infection. IBRV can cause latent infection in the spinal ganglion and trigeminal ganglion, and the infected cattle may have recessive infection, which is ignored because they do not show any clinical features, but the recessively infected cattle will continue to shed the virus to the outside world, causing other susceptible Infection in infected cattle. The same is true for recovered sick cattle. The virus is latent in the body. When the external environment chan...

Claims

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Application Information

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IPC IPC(8): C07K14/03C12N15/38A61K39/245A61P31/22A61P11/00A61P11/02
CPCA61K39/12A61P11/00A61P11/02A61P31/22C07K14/005C12N2710/16022C12N2710/16034
Inventor 曹文龙孔迪滕小锘张大鹤
Owner 苏州世诺生物技术有限公司
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