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IBV Vero cell adapted strain established on basis of reverse genetic technology

A reverse genetic technology, reverse genetic technology, applied to genetically modified cells, cells modified by introducing foreign genetic material, animal cells, etc., can solve the problems of cells that are not suitable for passage, and restrict the industrial production of vaccines, etc.

Active Publication Date: 2016-11-16
SICHUAN UNIV
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Problems solved by technology

However, virus strains such as IBV H120 can only be cultured through chicken embryos or primary cells and are not suitable for passage cells, which restricts the industrial production of vaccines

Method used

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  • IBV Vero cell adapted strain established on basis of reverse genetic technology

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Embodiment Construction

[0005] 1. The reverse genetic system of avian infectious bronchitis virus H120 strain used in the present invention was constructed by the Sichuan Key Laboratory of Animal Disease Prevention and Control and Food Safety of Sichuan University. The cell-adaptive strain Beaudette P65 strain S plasmid was donated by the professor of Nanyang Technological University, Singapore, and the passaged cell BHK21 used was preserved by the Sichuan Provincial Key Laboratory of Animal Disease Prevention and Control and Food Safety, Sichuan University. Anti-IBV serum was purchased from China Veterinary Drug Administration. Restriction enzymes BsaI, BsmB I, and NciI were purchased from NEB Company, high-fidelity PCR enzymes, pMD19-T, and T4 ligase were purchased from Dalian Bao Biological Co., Ltd., gel recovery kit was purchased from TIANGEN Co., Ltd., primers Designed and sequenced by Shanghai Sangon Co., Ltd.

[0006] 2. According to the No See'M technique, a pair of primers were designed to...

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Abstract

The invention discloses a poultry infectious bronchitis virus H120 reverse vaccine strain R-H120-Beaudette P65 (S) (Figure 1). The reverse vaccine strain is obtained by applying No See' M technology to replace S genes of an H120 strain with S genes of a PCR (polymerase chain reaction) amplification cell adapted strain Beaudette P65 strain. The reverse vaccine strain is adapted to Vero cells and can provide effective protecting force to chicken flocks. Through the reverse vaccine strain, the problem that IBV vaccine strains cannot be cultured in passage cells during production is solved, so that probability that vaccines are prone to being contaminated by exogenous viruses in the process of production is avoided, vaccine production quality is improved, and vaccine production cost is lowered greatly.

Description

technical field [0001] The invention relates to the field of animal medicine bioengineering, and relates to an avian infectious bronchitis virus H120 reverse vaccine strain R-H120-Beaudette P65 (S) adapted to Vero cells. Background technique [0002] Avian infectious bronchitis (IB) caused by avian infectious bronchitis virus (IBV) is one of the major infectious diseases that endanger the poultry industry. It is characterized by respiratory symptoms, a 30-50% drop in laying hen egg production, and a decline in egg quality. , renal lesions and gastrointestinal lesions are the main features. In my country, the disease incidence rate is 100%, the mortality rate is 10-30%, and the economic loss caused by the disease exceeds one billion yuan every year in China. The representative strain (H120) of avian infectious bronchitis virus (IBV) vaccine can only be cultivated through chicken embryos, which has problems such as high production cost, unstable production quality, and easy c...

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Application Information

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IPC IPC(8): C12N5/10A61K39/215A61P31/14
CPCA61K39/12A61K2039/53C12N5/0686C12N2510/00C12N2770/20034
Inventor 王红宁杨鑫周泷周英顺门帅
Owner SICHUAN UNIV
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