A method for infectious recombination cloning against type II type VII epidemic ndv strain dhn3

A technology of DHN3, cloning method, applied in the field of infectious recombinant cloning, can solve problems such as limitation and limitation

Active Publication Date: 2021-03-26
ZHAOQING INST OF BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the infectious cDNA clone of NDV has been established internationally, it is basically limited to the Latasa strain in China, and is basically limited to basic and clinical research levels. This may be related to the R&D technical capabilities of domestic vaccine production units

Method used

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  • A method for infectious recombination cloning against type II type VII epidemic ndv strain dhn3
  • A method for infectious recombination cloning against type II type VII epidemic ndv strain dhn3
  • A method for infectious recombination cloning against type II type VII epidemic ndv strain dhn3

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Experimental program
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Embodiment 1

[0067] 1. Main instruments and reagents

[0068] THZ-100 electric thermostatic incubator was purchased from Shanghai Yiheng Scientific Instrument Co., Ltd.

[0069] A17105653 clean bench was purchased from Suzhou Antai Air Technology Co., Ltd.

[0070] The DK-8D three-hole electric heating constant temperature water tank was purchased from Shanghai Yiheng Scientific Instrument Co., Ltd.

[0071] THZ-100 constant temperature incubator was purchased from Shanghai Yiheng Scientific Instrument Co., Ltd.

[0072] The BCD-579WE Haier refrigerator was purchased from Haier.

[0073] 5424R centrifuge; Eppendorf PCR instrument; Thermo1300 SERIES A2 biological safety cabinet, all purchased from Eppendorf.

[0074] DNA / RNA co-extraction kit (AP-MN-BF-VNA-250G) was purchased from AXYGEN, and TIAN prep Mini PlasmidKit (DP103-03) was purchased from Tiangen Co., Ltd. Gel Extraction Kit (D2500-02) was purchased from OMEGA; M-MLVRT (2641A) was purchased from TAKARA; RRI (2313A) was purchase...

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Abstract

The invention belongs to the technical field of veterinary biological products, and discloses a recombinant cloning method for the type II type VII popular NDV strain DHN3. The target fragments are NP gene, P gene and L gene respectively; Step 2: construct the DHN3 whole genome expression vector; recombine the DHN3 whole genome obtained by artificial recombination into the vector plasmid to obtain the whole genome expression vector; the sequence of the DHN3 whole genome is in sequence List SEQ ID NO 1; Step 3: co-transfect BHK-21 cells with three kinds of helper plasmids and DHN3 whole genome expression vector to obtain virus liquid containing recombinant virus rDHN3; NP gene is located in the sequence list SEQ ID NO: 1 at 1-1591nt; P gene is located at 1925-3109nt in SEQ ID NO: 1 of the sequence listing; L gene is located at 8166-15192nt in SEQ ID NO:1 of the sequence listing. This method can successfully clone a recombinant virus with the same original strain and no mutation.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to an infectious recombination cloning method for class II type VII epidemic NDV strain DHN3. Background technique [0002] Newcastle disease is a highly contagious and fatal disease caused by Newcastle disease virus (NDV) that mainly affects chickens, turkeys, wild birds and ornamental birds, commonly known as fowl plague. It is a highly contagious, acute and severe infectious disease. Humans occasionally become infected, manifesting as conjunctivitis. The World Health Organization (OIE) lists it as an infectious disease that must be notified, and the Ministry of Agriculture of my country also lists it as a class of animal diseases that must be notified. Because of its fast transmission speed and 100% morbidity and fatality rate, once it spreads, it will seriously harm the poultry industry and cause immeasurable losses. [0003] In addition to reasonable f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/45C12N15/85C12N15/66C12N7/01
CPCC07K14/005C12N7/00C12N15/66C12N15/85C12N2760/18121C12N2760/18122
Inventor 陈瑞爱黄梅王楠楠李延鹏刘定祥叶俊贤罗琼杨小云董楠
Owner ZHAOQING INST OF BIOTECHNOLOGY CO LTD
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