Method for expressing androctonus australis hector toxin by saccharomyces cerevisiae

Abstract

The invention relates to a method for expressing androctonus australis hector toxin by saccharomyces cerevisiae, successively comprising the following steps: providing a androctonus australis hector toxin gene AaHIT1; building a recombinant plasmid pYES2 / AaHIT1 expressing AaHIT1 protein in yeast cells; converting recombinant plasmid pYES2 / AaHIT1 into saccharomycetes cells; screening to obtain a converted bacterial strain AINVSC1 and identifying; cultivating the converted bacterial strain AINVSC1, and expressing androctonus australis hector toxin protein. The obtained androctonus australis hector toxin protein expressed by eukaryon has favourable insecticidal activity on armyworms and is suitable for producing in the biopesticide field.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a method for expressing North African scorpion toxin by Saccharomyces cerevisiae. Background technique [0002] Mythimna seperata Walker is an insect of the order Lepidoptera, also known as shaving insects and army insects. It is an important pest of food crops in my country. It migrates back and forth from north to south every year. When a large outbreak occurs, it can eat up the leaves of the crops, resulting in a large area of ​​production reduction. Or no harvest, and it has no diapause, high reproduction rate, and large body shape, which is representative to a certain extent. In China, many pesticide research institutions regard it as the target of new pesticide screening. [0003] At present, the control measures of armyworm mainly use chemical pesticides. However, the large-scale production and use of chemical pesticides has brought about three major problems: (1) env...

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Problems solved by technology

However, the large-scale production and use of chemical pesticides has brought about three major problems: (1) environmental pollution, increased residual toxicity, and both humans and animals have been poisoned; ecological balance

(4) The use of traditional chemical pesticides with high residues has caused the residues of agricultural and livestock products to seriously exceed the standard, resulting in blocked exports

[0006] The work of expressing North African scorpion toxin through prokaryotic system is in progress. The existing problem is that the activity of the expression product is low. The main reason is that AaHIT1 is a eukaryotic gene, and eukaryotic gene expression in prokaryotic system is prone to inclusion bodies, resulting in relatively low activity. low expression product

Saccharomyces cerevisiae is a probiotic closely related to people's daily life, and will not pose a threat to the environment; although researchers in this field can speculate that the North African scorpion toxin gene, as a eukaryotic gene, should be in the eukaryotic It has high activity when expressed in the expression system, but so far it has not been able to find a method to efficiently express North African scorpion toxin and use it to kill armyworms by using Saccharomyces cerevisiae

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