Hybridoma strain capable of secreting monoclonal antibodies for preventing tobacco mosaic virus and monoclonal antibody applications thereof

Abstract

The invention discloses a hybridoma strain capable of secreting monoclonal antibodies for preventing tobacco mosaic virus (TMV) and monoclonal antibody applications thereof. The purified TMV particles are taken as the antigen to immunize a BALB / c mouse, through the steps of cell fusion, screening, and cloning, a hybridoma strain 3A7, which can stably pass the characteristics to the next generation and secret monoclonal antibodies for preventing TMV, is obtained, and the preservation number of the hybridoma strain is CGMCC No.9338. The ascites indirect ELISA titer of monoclonal antibody secreted by the hybridoma strain can reach 10-8, and the antibody type and subtype are IgG1, kappa chain. The monoclonal antibody secreted by the hybridoma strain can carry out specific reactions with TMV. A dot-ELISA virus detection method is established on the basis of the 3A7 monoclonal antibody and is used to detect TMV in plants such as tobaccos, and the like. The hybridoma strain 3A7, the monoclonal antibody secreted by the hybridoma 3A7, and the establishment of related serum detection method provide technological and material support for the detection, diagnosis, and scientific prevention and control on TMV.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-tobacco mosaic virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Tobacco mosaic virus (TMV) is one of the more important viruses in plant viruses. It is also the type species of Tobamovirus. It is an RNA virus, the virion is rod-shaped, the length is 300-310 nm, and the diameter is 18 nm; the TMV genome is a single-stranded positive-sense RNA with a length of 6.3-6.6kb. The 5' end of the genome contains a cap structure. A non-coding region (UTR) of about 70 nucleotides; the UTR at the 3' end is 200-400 nt long and can be folded into a tRNA-like structure. The genome encodes at least four proteins, two proteins of 124-132 kDa and 181-189 kDa at the 5' end are related to virus replication, and the 181-189 kDa protein is produced by the read-through of the 124-132 kDa protein terminator. The ...

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Problems solved by technology

[0004] In order to grasp the incidence and hazards of tobacco mosaic virus (TMV) disease in my country, strengthen the early monitoring and early warning of tobacco mosaic virus disease in my country, and scientifically guide the prevention and control, it is urgent to establish a high-throughput detection method for the detection of tobacco mosaic virus disease. At present, there is no such high-throughput detection method, and only low-efficiency electron microscope observation, RT-PCR method, nucleic acid electrophoresis and other methods are used for small sample detection

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