Fungus expression vector and construction and screening method thereof

Abstract

The invention discloses a fungus expression vector. The expression vector is a constructive expression vector which consists of a plasmid expression vector pEX and an acidic trehalase gene serving as a screening marker, wherein the plasmid expression vector pEX is formed by removing a Bar gene in an expression vector pBarEx; and the screening marker gene belongs to acidic trehalase genes such as an ataxia telangiectasia mutated (ATM) gene or an ancient technology activation (ATA) gene. In the invention, a safe and environment-friendly screening marker is used for the fungus expression vector,and insect pathogen project fungi constructed by the fungus expression vector are released to the environment and cannot adversely influence the environment due to the screening marker. The fungus expression vector has a unique design principle, an ingenious construction method, a simple program, a simple screening program for an insect pathogen fungus transgenic bacterial strain of the fungus expression vector and high screening efficiency, does not influence the metabolic balance of the fungi, and has a potential and wide application prospect in the transgenic fungus research and marketization process.

Description

technical field [0001] The invention relates to a fungal expression vector and its construction method and screening method, in particular to a fungal expression vector using acid trehalase gene (Acid trehalase gene) as a screening marker and its construction and screening method, belonging to the field of biotechnology. Background technique [0002] Pest control is an important part of agricultural production. Chemical pesticides play an active role in controlling pests and increasing agricultural production. However, the long-term use of chemical pesticides has caused a series of environmental and food safety problems. Due to the advantages of safety and sustainable control, microbial biological control has been widely concerned by people. Insecticidal microbial pesticides are a large class of biological pesticides developed by using insect pathogenic microorganisms. Fungi are the largest group of insect pathogenic microorganisms. Under natural conditions, insects caused b...

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Problems solved by technology

[0004] However, at present, the screening markers for constructing engineering strains of insecticidal fungi at home and abroad use fungicide resistance markers including Benomyl resistance markers ( β-tubulin genes), herbicide resistance markers ( bar gene), hygromycin resistance marker ( hph Genes), genetically engineered bacteria containing these resistance selection markers (such as herbicide resistance markers bar Genetic engineering bacteria) enter the natural environment as biological pesticides. Once gene drift occurs, such as being absorbed by microorganisms such as weeds and plant pathogenic fungi in nature, weeds will become resistant to herbicides, and pathogenic microorganisms will develop drug resistance. , will have a great impact on the environment and ecosystem, so the selection of screening markers is an important factor restricting the wide application of engineering strains of insecticidal fungi

At present, no safe screening markers successfully used in fungal transformation at home and abroad have been reported.

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