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Sensitive cell line SLAM-MDCK of canine distemper virus and construction method and application of sensitive cell line SLAM-MDCK

A technology of canine distemper virus and sensitive cells, which is 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 disease progression

Active Publication Date: 2017-09-15
WUHAN KEQIAN BIOLOGY CO LTD
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  • Claims
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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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  • Sensitive cell line SLAM-MDCK of canine distemper virus and construction method and application of sensitive cell line SLAM-MDCK
  • Sensitive cell line SLAM-MDCK of canine distemper virus and construction method and application of sensitive cell line SLAM-MDCK
  • Sensitive cell line SLAM-MDCK of canine distemper virus and construction method and application of sensitive cell line SLAM-MDCK

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Embodiment Construction

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

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

[0069] 1. Cell and plasmid gene 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 retroviral vector pRetroX-Tet3G were purchased from Clontech.

[0070] 2. Commonly used biological reagents: DMEM medium was purchased from Gibco, fetal bovine serum was purchased from PAN Biotech, Germany, Phanta Super-Fidelity DNA Polymerase, DNA Maker DL2000, and DNA Maker DL5000 were purchased from Nanjing Novizan Biotechnology Co., Ltd. BamH Ⅰ, ECOR Ⅰ restriction endonuclease, dNTP MIX...

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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 Morbillivirus in the Paramyxoviridae family. The main clinical features are biphasic fever, mucous membrane inflammation of the eyes, nose, and digestive tract, and catarrhal pneumonia, dermatitis, and neurological symptoms. Since it was first reported in 1809, the disease has become a worldwide epidemic, seriously affecting companion animals and wildlife. CDV is divided into six types: Asia-I, Asia-II, Europe, America-II, Arctic and Vaccine genotype, but only one serotype is recognized. [0003] The hosts of natural infection of CDV are very extensive and present a trend of continuous expansion. It has expanded f...

Claims

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Application Information

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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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