Recombinant protein for pepper mottle virus and preparation method thereof, polyclonal antibody and preparation method for polyclonal antibody, detection test kit and application for detection test kit

Abstract

The invention discloses a recombinant protein for pepper mottle virus and a preparation method thereof, a polyclonal antibody and a preparation method for the polyclonal antibody, a detection test kitand an application for the detection test kit. The recombinant protein for the pepper mottle virus is a PepMoV CP protein and is obtained through the following steps: with a PepMoV CP gene as a template, carrying out PCR amplification on the PepMoV CP gene so as to obtain an amplification product, and carrying out enzyme digestion, connection, conversion and separation. The polyclonal antibody isobtained by immunizing white rabbits with the PepMoV CP recombinant protein as an immunogen. The detection test kit composed of the polyclonal antibody can be applied to serological detection of thepepper mottle virus. The method provided by the invention can realize rapid and high-throughput detection, and has the advantages of rapidness, simplicity, convenience, strong specificity, high sensitivity and reliable detection result. The detection test kit provided by the invention can be used for rapid detection, prediction and forecast of the pepper mottle virus in production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a recombinant protein of capsicum mottle virus and its preparation method, a polyclonal antibody and its preparation method, a detection kit and its application. Background technique [0002] pepper mottle virus ( Pepper mottle virus, PepMoV) is a member of the family Potatoviridae ( Potyviridae ) Potatovirus ( Potyvirus ), can be transmitted mechanically or by the vector aphid, and can infect a variety of important Solanaceae crops. Infestation of pepper usually results in yellowing of pepper leaves, and can also cause necrosis of leaf veins or necrosis of leaf buds or flower leaves in some pepper varieties (Isolation and characterization of pepper mottle virus infecting tomato in Korea. Plant Pathol J, 2008, 24(2): 152-158). PepMoV alone infects peppers, usually causing little yield loss; however, the symptoms of co-infection with plant viruses of Pepper mild mottle virus are...

AI Technical Summary

Problems solved by technology

PepMoV alone infects peppers, usually causing little yield loss; however, the symptoms of co-infection with plant viruses of Pepper mild mottle virus are more severe than those of these two viruses alone, and can Caused severe yield loss in pepper (Biological, physical and cytological properties of pepper mottlevirus-SNU1 and its RT-PCR detection. Plant Pathol J, 2006, 22(2): 155-160)

In 2016, the virus was detected on peppers in Hunan, Guangdong and Fujian provinces in my country, but the distribution, host range, infection cycle and harm of the virus in my country have not been reported so far. Therefore, the establishment of the virus The rapid detection technology system can provide technical means for monitoring the occurrence, distribution and harm of the virus, and can also provide a scientific basis for scientifically assessing the harm of the virus to agricultural production

[0003] The currently reported PepMoV detection method is mainly the RT-PCR method, which requires expensive instruments such as refrigerated high-speed centrifuges, PCR instruments, and gel imaging systems, as well as molecular biology reagents such as reverse transcriptase and high-fidelity Polymerase. At the same time, the detection process requires complex sample processing processes, such as total RNA extraction, reverse transcription; and the amplification process of more than 2 hours, and the tedious electrophoresis UV observation process

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