Co-immunoprecipitation method suitable for nosema bombycis

Abstract

The invention discloses a co-immunoprecipitation method suitable for nosema bombycis. The three technical problems in the prior art are solved, wherein as the first problem, spore total protein for a co-immunoprecipitation experiment is difficult to extract, and as the sporoderm of the nosema bombycis is thick, common protein extraction liquid cannot enter a spore, and protein cannot be fully extracted; as the second problem, due to the gravity, immune ball beads for co-immunoprecipitation can be naturally sunk to the bottom in the reacting process, stacked ball beads, protein liquid and antibodies cannot be fully combined, and the experiment result is influenced; as the third problem, most antibodies for the co-immunoprecipitation experiment are self-prepared polyclonal antibodies, the specific effect of the antibodies is poor, and nonspecific bands usually appear; in addition, detection to target protein can also be influenced by heavy chains and light chains of the antibodies. By means of the co-immunoprecipitation method suitable for the nosema bombycis, the specificity of the nosema-bombycis co-immunoprecipitation is improved, stability is high, and repeatability is good.

Description

technical field [0001] The invention belongs to the technical field of molecular biology experiments, and in particular relates to a method suitable for co-immunoprecipitation of the microsporidium silkworm. Background technique [0002] Nosema bombycis is the causative agent of silkworm microsporidia, which belongs to obligate intracellular parasitic eukaryotes. Because it can cause great harm to sericulture through vertical transmission, it is listed as a legal quarantine object for sericulture production by sericulture countries and regions. However, how to effectively solve the quarantine and prevention of microparticle diseases has become a difficult problem in current research. With the completion of the sequencing data of the silkworm Microsporidia genome, the study of Microsporidia has entered the field of molecular level, and some important results have been achieved. Septin and other gene family members are involved in the process of microsporidian adhesion and in...

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Problems solved by technology

[0004] However, this classic technique has deficiencies in the research on the interaction proteins of N. silkworm, mainly in three aspects: First, it is difficult to extract the total spore protein used in co-immunoprecipitation experiments. The spore wall itself is thick, and the ordinary protein extraction solution cannot penetrate deep into the spores, nor can it fully extract the protein; second, the immunoglobulin beads used for co-immunoprecipitation will naturally sink to the bottom during the reaction process due to gravity, As a result, the accumulated beads cannot be fully combined with the protein solution and antibodies, which affects the experimental results; third, most of the antibodies used in the co-immunoprecipitation experiments are self-made polyclonal antibodies, the specificity of the antibodies is not good, and non-specificity often occurs. In addition, the heavy and light chains of the antibody itself will also affect the detection of the target protein

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