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Canine distemper virus sensitive cell line slam-mdck and its construction method and application

A canine distemper virus and sensitive cell technology, applied in the field of cell lines, can solve the problems of complex methods and high laboratory requirements, and achieve the effects of high proliferation titer, good safety, and rapid lesions

Active Publication Date: 2021-01-15
WUHAN KEQIAN BIOLOGY CO LTD
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Problems solved by technology

However, the virus system constructed by Jiang Qian et al. is based on the HIV backbone and requires co-transfection of 3-4 plasmids. The method is relatively complicated and requires high laboratory requirements.

Method used

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  • Canine distemper virus sensitive cell line slam-mdck and its construction method and application
  • Canine distemper virus sensitive cell line slam-mdck and its construction method and application
  • Canine distemper virus sensitive cell line slam-mdck and its construction method and application

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Embodiment Construction

[0067] In order to better explain the present invention, the main content of the present invention is further illustrated below in conjunction with specific embodiments, but the content of the present invention is not limited to the following embodiments.

[0068] The main materials, reagents and instruments involved in the following specific implementation

[0069] 1. Cells and plasmid genes Canine kidney cell line (MDCK) was provided by ATCC, and GP2-293 cells were purchased from Clontech Company. The SLAM gene was synthesized by Shanghai Jierui Bioengineering Co., Ltd., and the pVSV-G plasmid and the retroviral vector pRetroX-Tet3G were purchased from Clontech Company.

[0070] 2. The commonly used biological reagents DMEM medium were purchased from Gibco company, fetal bovine serum was purchased from German PAN Biotech company, Phanta Super-Fidelity DNA Polymerase, DNA Maker DL2000, DNA Maker DL5000 were purchased from Nanjing Novizan Biotechnology Co., Ltd., BamH Ⅰ, ECOR...

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Abstract

The invention discloses a sensitive cell line SLAM-MDCK of a canine distemper virus and a construction method and application of the sensitive cell line SLAM-MDCK. A canine signaling lymphocytic activation molecule (SLAM) is constructed into an MDCK cell through a retroviral vector, and as a result the cell line SLAM-MDCK capable of stably expressing a canine SLAM gene is constructed. After being infected by the canine distemper virus, the cell line SLAM-MDCK is quick in lesion and high in viral titer, thereby being suitable for separation and culture of the canine distemper virus. Thus, the cell line SLAM-MDCK is of important significance to separation of the canine distemper virus, definite diagnosis of canine distemper virus infection as well as research, development and production of vaccines for the canine distemper virus.

Description

technical field [0001] The present invention relates to the field of cell lines, in particular to a canine distemper virus-sensitive cell line SLAM-MDCK and its construction method and application. Background technique [0002] Canine distemper (CD) is an acute, febrile and highly contagious infectious disease caused by Canine distemper virus (CDV) of the genus Measles virus in the Paramyxoviridae family. The main clinical features are bipolar fever, mucosal inflammation in the eyes, nose, and digestive tract, as well as catarrhal pneumonia, dermatitis, and neurological symptoms. Since it was first reported in 1809, the disease has shown a worldwide epidemic, with severe effects on companion and wildlife. CDV is divided into six different types: Asia-I, Asia-II, Europe, America-II, Arctic and Vaccine. genotype, but only one serotype is recognized. [0003] The natural infection hosts of CDV are very extensive and show an ever-expanding trend. It has been expanded from th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/867C12N15/66C12N15/12C12N5/10C12N7/00C12R1/93
CPCC12N5/0686C12N7/00C12N15/66C12N15/86C12N2510/00C12N2760/18451C12N2800/107
Inventor 韩丽李文珂张安定
Owner WUHAN KEQIAN BIOLOGY CO LTD
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