Serum IV-type avian adenovirus penton protein, preparation method and application thereof
A penton protein and poultry adenovirus technology, applied in the field of immunology, can solve the problems of easy amplification, etc., and achieve the effects of easy amplification, increased expression, and easy industrial scale production
Pending Publication Date: 2019-09-27
ZHAOQING DAHUANONG BIOLOGIC PHARMA +1
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Problems solved by technology
[0005] In view of the above problems, the present invention uses a baculovirus expression system for expression, which solves the problem of protein activity on the one hand, and on the other hand, sf9 cells are easy to scale up and culture in suspension, and are easy to produce on an industrial scale
Method used
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Embodiment 1
[0034] Example 1: Preparation of anti-group I serotype 4 avian adenovirus penton protein:
[0035] 1. Construction of recombinant Bacmid vector:
[0036] 1.1 Optimize the penton sequence:
[0037] Referring to the sequence of group I adenovirus serotype 4 (Genbank accession number: MH454598), the penton gene sequence was optimized for sf9 insect cell preferred codons, and the optimized penton gene sequence was obtained as shown in SEQ ID NO.1.
[0038] 1.2 Preparation of recombinant Bacmid vector:
[0039] Entrust Sangon Bioengineering (Shanghai) Co., Ltd. to synthesize the sequence of SEQ ID NO.1 (penton gene) and add a his tag to the N-terminal of the target gene penton;
[0040] Sangon Bioengineering (Shanghai) Co., Ltd. cloned the penton gene into pFastBac- 1 carrier, the obtained carrier pFastBac-1-penton (the carrier map is as follows figure 1 ).
[0041] Perform PCR verification on the vector pFastBac-1-penton, and prepare a 50 μL PCR reaction system with the fol...
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Abstract
The invention discloses a serum IV-type avian adenovirus penton protein, a preparation method and application thereof, and belongs to the field of amynology. The preparation method comprises the following steps: chemically synthesizing a penton gene sequence shown as SEQ ID NO.1; preparing a recombinant pFastBac-1-penton carrier; using the prepared recombinant pFastBac-1-penton carrier to transfect an sf9 cell, and obtaining a recombinant protein through expression of the sf9 cell; and carrying out Ni-NTA purification on the obtained recombinant protein to obtain purified serum IV-type avian adenovirus penton protein. The serum IV-type avian adenovirus penton protein is obtained by the preparation method. The invention discloses application of the serum IV-type avian adenovirus penton protein in the preparation of vaccines for preventing avian adenovirus. The serum IV-type avian adenovirus penton protein has the advantages that gene synthesis does not need a template, and is more economic, simpler and more convenient; the expression quantity of penton can be improved preferably; and the protein is easy for industrial scaled production.
Description
technical field [0001] The invention belongs to the field of immunology, and in particular relates to a serum type 4 avian adenovirus penton protein, a preparation method and application thereof. Background technique [0002] Group I serotype 4 avian adenovirus can cause chicken pericardial effusion hepatitis syndrome, which is an acute infectious disease affecting chickens. The main lesion is clear or light yellow watery or jelly-like liquid in the pericardial cavity, liver enlargement, Partial necrosis or discoloration may even have inclusion bodies, which can be transmitted horizontally or vertically, with high morbidity and mortality. Pericardial effusion hepatitis syndrome caused by group I serotype 4 avian adenovirus was first reported in Pakistan in 1987. It appeared in my country in 2012, became widespread in 2014, and spread to the whole country in 2015-2016. At present, the incidence of the disease in chicken flocks in my country is relatively high, and it is grad...
Claims
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Login to View More IPC IPC(8): C12N15/85C07K14/075A61K39/235A61P31/20
CPCC12N15/85C07K14/005A61K39/12A61P31/20C12N2800/105C12N2800/22C12N2710/10251C12N2710/10222C12N2710/10234A61K2039/552
Inventor 任广彩陈瑞爱黄妙容李延鹏罗琼张文炎刘之文
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA