Cell line for stable expression of peste des petits ruminants virus receptor Nectin-4 and construction method of cell line

A technology of Peste des petits ruminants, mdbk-nectin-4, applied in the direction of receptors/cell surface antigens/cell surface determinants, viruses/phages, cells modified by introducing foreign genetic material, etc., can solve the problem of direct transfection Low efficiency and other issues, to achieve good genetic stability

Inactive Publication Date: 2018-03-16
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

In March 2014, many suspected PPR epidemics occurred in Inner Mongolia, Liaoning Province, Hunan Province, Jiangxi Province, Jiangsu Province and Anhui Province, which were confirmed as PPR epidemics by the National Research Center for Exotic Animal Diseases. Alarm bells are ringing! As there is currently no specific tre

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  • Cell line for stable expression of peste des petits ruminants virus receptor Nectin-4 and construction method of cell line
  • Cell line for stable expression of peste des petits ruminants virus receptor Nectin-4 and construction method of cell line
  • Cell line for stable expression of peste des petits ruminants virus receptor Nectin-4 and construction method of cell line

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[0035] In order to make the technical means realized by the present invention, creative features, goals and effects easy to understand, the following in conjunction with specific embodiments and appended Figure 1~5 To further illustrate the present invention, the content mentioned in the embodiment is not to limit the present invention. If no specific conditions are indicated in the examples, it shall be carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used are commercially available routine products, if the manufacturers are not indicated.

[0036] 1. Construction of pLOV-eGFP-Nectin-4 expression vector

[0037] The Nectin-4 gene was synthesized according to the sheep Nectin-4 gene sequence published by Genbank (Accession No. JX944709).

[0038] The Nectin-4 gene sequence (SEQ ID NO.3): (accession number JX944709)

[0039] atgcctctgtccctgggagccgagatgtggggtcctgtggcctggctgctgctgctgctcctggcatcatt...

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Abstract

The invention discloses a cell line for stable expression of a peste des petits ruminants virus receptor Nectin-4 and a construction method of the cell line. According to the invention, by virtue of asynthetic Nectin-4 gene and on the basis of a lentiviral vector system, a recombinant plasmid pLOV-eGFP-Nectin-4 is constructed. The recombinant plasmid, together with pSPAX2 and pMD2.G plasmid, is applied to co-transfection of a 293T cell, so that retrovirus-like particles are obtained. A supernatant-infected MDBK cell is cultivated by virtue of cells containing the retrovirus-like particles, and puromycin screening and purifying are conducted, so that the MDBK cell for stable expression of Nectin-4 protein is obtained, and the MDBK cell is named as MDBK-Nectin-4. Through the construction ofthe cell line, an experimental material is provided for deep research of an interaction between PPRV and the receptor Nectin-4 and for the clinical rapid isolation of the PPRV is provided.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a cell line stably expressing Peste des Petits Ruminants virus receptor Nectin-4 and a construction method thereof. Background technique [0002] Peste des petits ruminants (PPR) is an acute, highly contagious infectious disease caused by Peste des petits ruminants virus (PPRV), which mainly infects small ruminants such as sheep and goats. The World Organization for Animal Health defines it as a disease that must be reported, and my country lists it as a first-class animal disease. The disease causes acute morbidity and mortality in animals as high as 100% and 90%, respectively. In recent years, PPR has shown a tendency to spread, and countries such as India and Pakistan around my country have experienced large-scale outbreaks. [0003] In July 2007, the PPR epidemic occurred for the first time in my country's Tibet region, and was quickly extinguished by relevant departmen...

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Application Information

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IPC IPC(8): C12N5/10C12N15/867
CPCC07K14/70503C12N15/86C12N2740/15043
Inventor 王勇王元红杨侃侃殷冬冬俞赵荣蒋书东
Owner ANHUI AGRICULTURAL UNIVERSITY
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