Preparation method of rice dwarf virus antibody

A dwarf virus and rice technology, which is applied in the field of virus antibody preparation, can solve the problem that the large molecular target protein is not easy to induce rice dwarf virus antibody, and achieve the effect of solving the problem of not being easy to induce expression and solving technical bottlenecks

Inactive Publication Date: 2017-01-11
刘铮
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Problems solved by technology

[0004] The present invention aims at the above problems and provides a method for preparing rice dwa

Method used

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Embodiment Construction

[0017] The present invention includes the following steps.

[0018] 1) Positive strains were inoculated in two tubes of LB liquid medium containing ampicillin and chloramphenicol at a ratio of 1:100, shaken for 3-4 hours, induced for 4-5 hours, and no IPTG was added to the other test tube as a control; take the bacteria Bacteria were collected and separated by gel electrophoresis and placed in Coomassie brilliant blue staining solution.

[0019] 2) Induce 100ml of bacterial liquid, collect the bacterial cells, conduct gel electrophoresis for the target protein, and pre-cool the gel after electrophoresis to obtain milky white protein bands, cut and recover the target protein bands with a blade, add Freund's adjuvant and grind until Emulsified, save for future use.

[0020] 3) Extract the total protein of SRBSDV-infected rice diseased plants and healthy rice plants, separate by gel electrophoresis, transfer to PVDF membrane, seal on a shaker at room temperature for 1 hour, and ...

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Abstract

The invention belongs to the technical field of virus antibody preparation, and particularly relates to a preparation method of a rice dwarf virus antibody. The invention provides the preparation method of the rice dwarf virus antibody, which solves the problem that macromolecular target proteins are generally not easy to induce. The preparation method comprises the following steps: (1) inoculating a positive strain in two tubes of LB fluid mediums containing penbritin and chloramphenicol according to a ratio of 1 to 100, shaking for 3 to 4 h, and performing induced culture for 4 to 5 h, wherein IPTG is not added in another test tube for contrast; taking bacterial fluid, collecting thalli, and putting into Coomassie brilliant blue staining fluid after the thalli are subjected to gel electrophoresis separation; and (2) inducing 100 ml of the bacterial fluid, collecting thalli, performing gel electrophoresis on target proteins, performing KCL pre-cooling on gel subjected to electrophoresis to obtain a milk white protein band, cutting and recycling the target protein band with a blade, adding the freund's adjuvant, grinding to obtain emulsified proteins, and storing the emulsified proteins for later use.

Description

technical field [0001] The invention belongs to the technical field of virus antibody preparation, and in particular relates to a preparation method of rice dwarf virus antibody. Background technique [0002] Southern rice black-streaked dwarf disease was first discovered in Yangjiang City, Guangdong Province in 2001, and gradually spread to neighboring provinces in the following years. In 2009, a large-scale outbreak occurred in 9 provinces in southern China, and the affected area was about 300,000 hm2. In the same year, the disease commonly occurred in 19 provinces in northern Vietnam. In 2010, more than 1.3 million hectares of rice were damaged in 13 provinces in southern China, and about 60,000 hectares of rice were damaged in 29 provinces in central and northern Vietnam. In 2010, the disease was also found in Kyushu, Japan. In addition, the disease can not only harm rice, but also maize. In 2009, the disease was also detected in maize in Shandong, Guangdong, Guangxi a...

Claims

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Application Information

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IPC IPC(8): C07K16/10
Inventor 刘铮
Owner 刘铮
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