Method for establishing interaction between phytoplasma and tubulin through co-immunoprecipitation

Abstract

The invention discloses a method for establishing interaction between phytoplasma and tubulin through co-immunoprecipitation. The target protein is microtubulin, the phytoplasma is wheat blue dwarf phytoplasma, and the mediator is Isaria calycina; the method comprises the following steps: full-length cloning of a mediator target protein gene, verification of interaction between a phytoplasma immune membrane protein and the mediator target protein through co-immunoprecipitation, co-localization of the phytoplasma immune membrane protein and the mediator target protein in Sf9 insect cells, yeast two-hybrid reinspection, and determination of interaction between the phytoplasma and microtubulin. The invention finds that the wheat blue dwarf phytoplasma and the heterosigmaea microtubulin generate interaction, the method avoids the situation that false positive occurs when the interaction between the phytoplasma and the vector target protein is verified, and the position of the interaction between the phytoplasma and the vector target protein is positioned.

Description

technical field [0001] The invention belongs to the technical field of plant pathology, and in particular relates to a method for establishing the interaction between phytoplasma and tubulin through co-immunoprecipitation. Background technique [0002] Phytoplasma is a single-celled prokaryote that depends on vector insects for transmission and obligately parasitizes in the phloem of plants. It has no cell wall and belongs to Class Mollicutes, phylum Tenericutes. Usually, when a new phytoplasma disease is discovered, the identification of the transmission vector is mainly carried out on the insect species found near the diseased plants. Insects with organs are tested to determine whether they contain phytoplasma in their bodies. In the early stage, the phytoplasma in insect media was observed by electron microscope, and later with the development of PCR technology, the poisonous insects were determined by extracting insect DNA. [0003] Wheat blue dwarf disease (WBD) is on...

AI Technical Summary

Problems solved by technology

At present, there is no report on the interaction between phytoplasma and tubulin protein (tubulin). In the identification method of the interaction protein, the commonly used yeast two-hybrid experiment is used to verify the interaction between phytoplasma and tubulin, which is prone to false positives. Therefore, to find out whether the immune membrane protein (IMP) of wheat blue dwarf phytoplasma and the microtubulin (Tubulin) of the leafhopper has an interaction is the key to ascertain the interaction mechanism between phytoplasma and mediator

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