Expression of cymbidium mosaic virus coat protein gene and preparation method for antibody of cymbidium mosaic virus coat protein gene

A coat protein and technology for building orchid leaves, applied in antiviral immunoglobulin, botany equipment and methods, biochemical equipment and methods, etc., can solve orchid ornamental value and economic value decline, orchid planting loss, plant growth No defects and other problems, to achieve the effect of high sensitivity, good quality, high protein content

Inactive Publication Date: 2010-06-23
ZHEJIANG UNIV
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Problems solved by technology

Studies have found that Jianlan mosaic virus often forms co-infection with Odontoglossum ringspot virus (ORSV), causing orchid leaves to form irregular chlorotic stripes, round spots, ring spots and even necrosis, resulting in poor plant growth, The flowers are small and few, and the flowering period is shortened, which greatl...

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  • Expression of cymbidium mosaic virus coat protein gene and preparation method for antibody of cymbidium mosaic virus coat protein gene
  • Expression of cymbidium mosaic virus coat protein gene and preparation method for antibody of cymbidium mosaic virus coat protein gene
  • Expression of cymbidium mosaic virus coat protein gene and preparation method for antibody of cymbidium mosaic virus coat protein gene

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Embodiment Construction

[0025] The invention prepares the shell protein of Jianlan mosaic virus with high content and good purity, and prepares CymMV antibodies with strong specificity, high sensitivity and good quality, and the immunocapture RT-PCR and ELISA methods established by using the prepared antibodies can be highly specific, Accurate, sensitive and rapid detection of CymMV can provide strong technical and product support for virus-free identification and disease-resistant breeding of orchids, and provide direct theoretical basis and technical support for the comprehensive prevention and treatment of this virus disease. It can also be applied to import and export inspections, Quarantine etc.

[0026] 1. Cloning of CymMV coat protein gene

[0027] Design primers based on the full-length sequence of the gene encoding CymMV coat protein in GenBank, upstream primers

[0028] CP-F is 5′-CG GGATCC ATGGGAGAGCCCACTCCA-3′, downstream primer CP-R is 5′-CG GTC GAC TTATTCAGTAGGGGGTGCAG-3′, the BamH ...

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Abstract

The invention discloses an expression of cymbidium mosaic virus (Cymbidium Mosaic Virus, CymMV) coat protein gene and a preparation method for antibody of cymbidium mosaic virus coat protein gene. A prokaryotic expression system is used for expressing the coat protein of the cymbidium mosaic virus, and the cymbidium mosaic virus coat protein gene immune rabbit, the BALB / C mouse are used for preparing the polyclonal anti-serum; and the hybridoma technique is used for preparing the monoclonal antibody, and the prepared antibody is used for establishing the immune capture RT-PCR and ELISA method for detecting the high specificity, sensitivity and accuracy of CymMV. The invention provides technical support for the diagnosis of the virus, disease-resistance breeding and cultivation of virus-free seedlings. The expression of cymbidium mosaic virus coat protein gene and the preparation method for its antibody provided by the invention provides material base and technical support for rapid detection and diagnosis of the cymbidium mosaic virus, typing and the study on molecular biology, and lays the foundations for prevention and treatment of cymbidium mosaic virus disease.

Description

technical field [0001] The field of the invention belongs to the field of biotechnology, and in particular relates to the expression of the coat protein gene of Jianlan mosaic virus and its antibody preparation method. Background technique [0002] Cymbidium mosaic virus (CymMV) is one of the main viruses that seriously damage orchids all over the world. The Jianlan mosaic virus particle is linear, with a size of about 475nm×13nm or 490nm×13nm, and the CymMV genome nucleic acid is a linear positive-strand ssRNA. Studies have found that Jianlan mosaic virus often forms co-infection with Odontoglossum ringspot virus (ORSV), causing orchid leaves to form irregular chlorotic stripes, round spots, ring spots and even necrosis, resulting in poor plant growth, The flowers are small and few, and the flowering period is shortened, which greatly reduces the ornamental value and economic value of orchids. In recent years, with the development of the orchid industry, the virus has sprea...

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Application Information

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IPC IPC(8): C12N15/40C12N15/70C07K14/08C07K16/10G01N33/569C12Q1/70C12Q1/68
Inventor 吴建祥洪健孟春梅
Owner ZHEJIANG UNIV
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