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Preparation method of duck tembusu virus infectious cDNA and preparation method of recombinant virus rDTMV-QY21

A duck Tembusu virus, infectious technology, applied in the direction of recombinant DNA technology, microorganism-based methods, botanical equipment and methods, etc., can solve the problems of heavy economic burden and unsatisfactory protection titer in the duck industry.

Pending Publication Date: 2022-03-18
岭南现代农业科学与技术广东省实验室肇庆分中心 +1
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Problems solved by technology

The protective potency of the vaccine is not so satisfactory that repeated vaccination is required, thus causing a heavy economic burden on the duck industry

Method used

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  • Preparation method of duck tembusu virus infectious cDNA and preparation method of recombinant virus rDTMV-QY21
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  • Preparation method of duck tembusu virus infectious cDNA and preparation method of recombinant virus rDTMV-QY21

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Experimental program
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Embodiment 1

[0054] 1. Experimental materials

[0055] Main equipment

[0056]5424R centrifuge; Eppendorf PCR instrument; Thermo1300 SERIES A2 biological safety cabinet, all purchased from Eppendorf. Electric constant temperature incubator HZ-100 (Yiheng Scientific Instrument Co., Ltd., Shanghai, China); three-hole electric heating constant temperature water tank DK-8D (Yiheng Scientific Instrument Co., Ltd., Shanghai, China); Haier BCD-579WE refrigerator (Haier, Shanghai, China) ; CO2 constant temperature incubator Forma371 (Thermo Company, the United States); ultra-clean bench SW-CJ-2FD (Suzhou Antai Air Technology Co., Ltd., Jiangsu, China); biological safety cabinet 1300SERIESA2 (Thermo Company, the United States); inverted optical microscope (Nikon company, Japan); high-speed centrifuge Centrifuge5804R (Eppendorf Company, Germany); pipette Researchplus (Eppendorf Company, Germany); PCR instrument C1000Touch (Bio-Rad Company, U.S.); electrophoresis instrument PowerPacBasic (Bio-Rad Co...

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Abstract

The invention belongs to the field of veterinary products, and discloses a preparation method of duck tembusu virus infectious cDNA (complementary deoxyribonucleic acid), which comprises the following steps: step 1, constructing subgenome plasmids, the subgenome plasmids are plasmids pF1, pF2, pF3 and pF4, and the plasmids pF1, pF2, pF3 and pF4 respectively contain target fragments F1, F2, F3 and F4; 2, obtaining target fragments: obtaining the target fragments f1, f2, f3 and f4 for in-vitro connection from the plasmids pF1, pF2, pF3 and pF4; and step 3, connecting target fragments f1, f2, f3 and f4 in vitro to obtain the duck tembusu virus infectious cDNA. The method can successfully clone the recombinant virus which is completely the same as the original virus strain and has no mutation.

Description

technical field [0001] The invention relates to the field of veterinary products, in particular to a method for preparing duck Tembusu virus infectious cDNA and a method for preparing recombinant virus rDTMUV-QY21. Background technique [0002] In April 2010, a new disease broke out in many duck farms across China. The characteristic symptoms of the disease include loss of appetite, decreased egg production, ovarian congestion, hemorrhage, degeneration, degeneration, and lymphocyte infiltration, and severe cases can cause death. After virus isolation and identification, it was determined that the pathogen causing the disease was a new flavivirus named Duck Tembusu Virus (DTMUV). In recent years, duck Tembusu virus disease, which is characterized by growth retardation and decreased egg production, has spread widely in duck farming areas in China, and has caused huge economic losses to China's waterfowl farming industry. However, the vaccines against the virus currently on t...

Claims

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Application Information

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IPC IPC(8): C12N15/40C12N15/85C12N7/01C12R1/93
CPCC07K14/005C12N7/00C12N15/85C12N2770/24021C12N2770/24022
Inventor 陈瑞爱王婷黄梅向程威杨洁杨泽坤
Owner 岭南现代农业科学与技术广东省实验室肇庆分中心
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