Artificially synthesized mcry1ba and mcry1ia gene sequences and gene combinations with high toxicity to lepidopteran pests

Abstract

The invention discloses artificially synthesized gene sequences of mcry1Ba and mcry1Ia with high toxicity to lepidoptera pests and a gene combination, and belongs to the technical field of biological prevention and treatment. According to the two new DNA sequences (shown by SEQ ID NO1 and SEQ ID NO2 in the sequence table) which are artificially optimized, transformed and synthesized by gene Bt cry1Ba and gene Bt cry1Ia which are high in toxicity to the lepidoptera pests, the conversion success rate of the gene Bt cry1Ba and the gene Bt cry1Ia in a transgenosis process is increased by using a codon sequence preferred by plants, and also the expression amount of the genes in transgenic plants and the stable expression characteristic are improved; by the combination application of the two genes, the insecticidal activity of the genes is improved. A new plant expression vector is constructed by using promoters for constitutive expression, so that the conversion rate is high, the expression amount is high and stable, and the artificially synthesized gene sequences can be used for researching the transgenic plants.

Description

technical field [0001] The invention belongs to the technical field of biological control, in particular to artificially synthesized mcry1Ba and mcry1Ia gene sequences and gene combinations with high toxicity to lepidopteran pests. Background technique [0002] Insect pests are an important factor in the reduction of crop production in the world. On average, about 10% of the total grain output is lost every year, and the direct economic loss reaches more than 10 billion US dollars. In the past few decades, the control has mainly relied on chemical pesticides. While making great contributions to agricultural production, chemical pesticides have caused serious consequences such as environmental pollution, human and animal poisoning, and ecological imbalance. In the face of these huge costs, the world is looking for and developing new pest control strategies and technologies. [0003] Bacillus thuringiensis (Bt) is a widely distributed Gram-positive bacteria. While forming sp...

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Problems solved by technology

However, in transgenic research, it has been found that the insecticidal protein gene derived from Bt is directly transferred into plants, and there are defects such as unstable expression products and low expression levels (van Aarssen et al, 1995, Plant Molecular Biology, 28:513-524 )

Specific problems include: 1) the natural Bt gene contains high AT, more than 60%, and the mRNA expressed by such a gene in the plant is very easy to be degraded by the plant; 2) there are intron cut points similar to eukaryotic genes in the natural Bt gene, Transcription terminator sequence, resulting in incomplete transcription, abnormal splicing of mRNA, etc.; 3) The codons used in natural Bt genes are quite different from those in plants, which will reduce the efficiency of protein translation; 4) Natural Bt genes are used as prokaryotic sources The structure of the gene is significantly different from that of plants and other eukaryotes, such as eukaryotes containing 5'-UTR sequence and polyA tail sequence at the 3' end

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