Recombinant baculovirus expressing novel duck reovirus outer capsid proteins and construction method thereof

A technology of recombinant baculovirus and construction method, which is applied in the field of recombinant baculovirus and its construction of new duck reovirus outer capsid protein, which can solve the problems of prokaryotic expression, protein expression, modification and processing, limited antigenic protein, etc.

Inactive Publication Date: 2014-03-19
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is: NDRV outer capsid proteins σB and σC are both antigenic proteins, which can stimulate the body to produce protective neutralizing antibodies, σB is a group-specific antigen, σC is a type-specific antigen, and obtained by purifying the virus The antigenic protein of the virus is very limited, and prokaryotic expression cannot modify and process the protein expression, and there is a certain gap between the expression product and the natural protein. Therefore, it is necessary to use the eukaryotic expression system to efficiently recombine and express the viral protein in vitro, and then prepare the NDRV gene. Engineering vaccines and conducting research on antiviral mechanisms are not new and effective ways

Method used

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  • Recombinant baculovirus expressing novel duck reovirus outer capsid proteins and construction method thereof
  • Recombinant baculovirus expressing novel duck reovirus outer capsid proteins and construction method thereof
  • Recombinant baculovirus expressing novel duck reovirus outer capsid proteins and construction method thereof

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Embodiment Construction

[0042] The present invention will be further described below in conjunction with example, but does not limit the present invention.

[0043] 1. Cells and virus strains

[0044] Transfection of Recombinant NDRV Capsid Protein Baculovirus Plasmid Using Sf9 Insect Cell Culture

[0045] 1) Sf9 cell culture medium is SF-900II SFM (Invitrogen Company, Cat. No. 10902-088)

[0046] 2) The cell culture method is as follows: take sf9 cells stored in liquid nitrogen, melt them quickly in a 37°C water bath, transfer them to a cell culture bottle in an ultra-clean bench, add an appropriate amount of medium, put them in a 27°C incubator, and cultivate for 2 to 3 days. Day, after the bottom of the culture bottle was covered, subculture was carried out.

[0047] 3) NDRV strains are 11 strains from Taihu Lake (Chen Zongyan, Zhu Yingqi, Wang Shichuan, Li Chuanfeng, Li Lu, Wang Bingbin, Fu Yuzhi, Gao Jingpeng, Wang Chao, Liu Guangqing. Isolation of a new duck-derived reovirus (TH11 strain) an...

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Abstract

The invention relates to a recombinant baculovirus expressing novel duck reovirus NDRV outer capsid proteins. The recombinant baculovirus is vAcNDRV-[sigma]B / [sigma]C, and co-expresses the NDRV outer capsid protein [sigma]B and the NDRV outer capsid protein [sigma]C. The invention also relates to a construction method of the recombinant baculovirus. The construction method includes performing RT-PCR amplification to respectively obtain specific gene fragments of the [sigma]B and the [sigma]C; obtaining a co-expression carrier pFbNDRV-[sigma]B / [sigma]C through gene cloning and screening; transforming a DH10Bac cell with the screened recombinant co-expression carrier to obtain a transformed AcNDRV-[sigma]B / [sigma]C Bacmid; and transfecting the transformed AcNDRV-[sigma]B / [sigma]C Bacmid to an sf9 insect cell to obtain a recombinant virus vAcNDRV-[sigma]B / [sigma]C in the sf9 insect cell.

Description

technical field [0001] The invention relates to the field of virology, in particular to a recombinant baculovirus of a novel duck reovirus outer capsid protein and a construction method thereof. The virus strain used in the present invention is Novel duck reovirus (NDRV for short) Anhui Taihu 11 strain. Background technique [0002] Duck hemorrhagic necrotizing hepatitis is a kind of duck reovirus (Novel duck reovirus, NDRV) caused by the new type of duck reovirus (NDRV) in the family Reoviridae. It is characterized by irregular liver necrosis and hemorrhage. The main characteristic of the blight. Different breeds of ducks are susceptible, and the younger the age or the higher the morbidity and mortality when they are concurrently infected, the majority are 5 to 10 days old, the course of disease is 5 to 7 days, the incidence rate is 5% to 20%, and the mortality rate is 2% to 15 days. %. [0003] Since 2005, Sichuan, Guangdong, Anhui and other provinces have successively ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85A61K35/76A61P31/12A61K39/15A61P31/14A61K35/765
Inventor 刘光清陈宗艳毕庄莉朱音奇李传峰孟春春
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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