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Newcastle disease virus vaccine

A technology for Newcastle disease virus and vaccine, applied in the direction of virus, antiviral immunoglobulin, virus peptide, etc., can solve the problems of high epidemic prevention cost, high cost, poor protection effect, etc., and achieve cost reduction, simplify the preparation process, The effect of good cross protection

Active Publication Date: 2022-05-31
天康制药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current Newcastle disease virus vaccines mainly have the following forms: 1. Chicken embryos produce inactivated vaccines, which have the disadvantages of complicated process, environmental pollution and high cost; 2. Recombinant expression of pathogenic related proteins using molecular biology techniques to prepare subunit vaccines The disadvantage is that the operation is cumbersome and the protection ability is poor
[0003] In China, the prevailing genotypes of Newcastle disease virus are mainly genotype II, genotype VII, or both subtypes. However, the current technology is not effective for the cross-protection of the two subtypes, and the cost of epidemic prevention is high. This puts a lot of pressure on clinical prevention and control

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0081] Type VII F protein SEQ ID NO.1 was constructed to obtain CHO-F (VII); type II HN protein SEQ ID NO.2 was constructed to obtain CHO-HN (II); two proteins were mixed to prepare a vaccine to obtain vaccine 1.

[0082] Downloaded from Geenbank the sequences of the Newcastle disease type VII F gene and type II HN gene sequences that are currently prevalent in the past five years, and compared and analyzed them. The codons were optimized and modified respectively to obtain SEQ ID NO.1 and SEQ ID NO.2 respectively. Both genes have a high degree of broad-spectrum and can increase the expression levels of the target proteins F and HN. F and HN genes were artificially synthesized by designing enzyme cutting sites.

[0083] The above-mentioned nucleotide sequences encoding F and HN proteins (with a His tag at the C-terminal), F through NotI and EcoRI sites, and HN through KpnI and XhoI sites were respectively inserted and cloned into the eukaryotic transfer vector pcDNA3.1. Use T...

Embodiment 2

[0094] Type II F protein SEQ ID NO.3 was constructed to obtain CHO-F (II); type VII HN protein SEQ ID NO.4 was constructed to obtain CHO-HN (VII); the obtained two proteins were mixed to prepare a vaccine to obtain vaccine 2.

[0095] Downloaded from Geenbank the sequences of the Newcastle disease type II F gene and VII HN gene sequences that have been prevalent in the past five years and compared them. The gene sequence was optimized and modified to obtain SEQ ID NO.3 and SEQ ID NO.4 respectively, which made the gene highly broad-spectrum and increased the expression levels of the target proteins F and HN. F and HN genes were artificially synthesized by designing enzyme cutting sites.

[0096] The above-mentioned nucleotide sequences encoding F and HN proteins (the C-terminal has a His tag), F through the XbaI and XhoI sites, and HN through the KpnI and XhoI sites, were inserted and cloned into the eukaryotic transfer vector pcDNA3.1. Use T4 DNA ligase to ligate overnight at...

Embodiment 3

[0106] Type II F protein and Type VII HN protein, fusion gene SEQ ID NO.6, was constructed to obtain CHO-F(II)-HN(VII); the fusion protein was used to prepare a vaccine, and vaccine 3 was obtained.

[0107] Downloaded from Geenbank the sequences of the Newcastle disease type II F gene and VII HN gene sequences that have been prevalent in the past five years and compared them. The gene sequence was optimized and modified to make the gene highly broad-spectrum, and the Newcastle disease virus was used to express the protein by fusing F and HN naturally through a linker (GAA). The F+HN gene was synthesized by dividing the labor by designing the enzyme cutting site.

[0108] The above nucleotide sequence encoding F+HN protein (the C-terminal has a His tag), F+HN was cloned into the eukaryotic transfer vector pcDNA3.1 by inserting HindIII and SmaI. Use T4 DNA ligase to ligate overnight at 16°C to obtain ligation products, transform Escherichia coli competent DH5a, spread on LB pla...

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Abstract

The invention relates to the field of biotechnology, in particular, it provides a Newcastle disease virus vaccine. Prepare the protein of Newcastle disease virus vaccine, including the F protein of Newcastle disease virus and the HN protein of Newcastle disease virus, wherein, the nucleotide sequence of encoding F protein is as SEQ ID NO.1 (gene type VII) or SEQ ID NO.3 ( Gene Type II), the nucleotide sequence encoding HN protein is shown in SEQ ID NO.2 (Gene Type II) or SEQ ID NO.4 (Gene Type VII). The protein is suitable for the eukaryotic cell expression system, simplifies the process for preparing the active ingredient of the Newcastle disease virus vaccine, and reduces the cost at the same time. Moreover, the protein has broad-spectrum antigenicity and can achieve a good cross-protection effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Newcastle disease virus vaccine. Background technique [0002] Newcastle Disease Virus (NDV) belongs to Paramyxoviridae, a single-stranded negative-sense RNA virus of the genus Mumps virus, and is one of the most important infectious viruses that endanger the poultry industry. The current Newcastle disease virus vaccines mainly have the following forms: 1. Chicken embryos produce inactivated vaccines, which have the disadvantages of complicated process, environmental pollution and high cost; 2. Recombinant expression of pathogenic related proteins using molecular biology techniques to prepare subunit vaccines The disadvantage is that the operation is cumbersome and the protection ability is poor. [0003] In China, the prevailing genotypes of Newcastle disease virus are mainly genotype Ⅱ, genotype Ⅶ, or both subtypes. However, the current technology is not effective for the cross-...

Claims

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Application Information

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IPC IPC(8): C07K14/125C07K19/00C07K16/10C12N15/45C12N15/85G01N33/569A61K39/17A61P31/14
CPCC07K14/005C07K16/1027C12N15/85G01N33/56983A61K39/12A61P31/14C12N2760/18122C12N2760/18134C07K2319/00G01N2333/125Y02A50/30
Inventor 贺笋李俊辉潘毅平张伟程兰玲潘晓梅毛丽萍王雨朦
Owner 天康制药股份有限公司
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