Bivalent recombination beauveria bassiana capable of killing mosquito larvae and preparation method of bivalent recombining beauveria bassiana

Abstract

The invention relates to bivalent recombination beauveria bassiana capable of killing mosquito larvae. Chromosome of the recombination beauveria bassiana is recombined with a scorpion neurotoxin gene AaIT1 and an agritol insecticidal crystal protein gene Cyt2B alpha; the beauveria bassiana is a bacterial strain of wild type beauveria bassiana; the sequence of the scorpion neurotoxin gene AaIT1 is shown in SEQ NO.2; and the sequence of the agritol insecticidal crystal protein gene Cyt2B alpha is shown in SEQ NO.5. A preparation method of the bivalent recombination beauveria bassiana comprises the following steps of optimizing the scorpion neurotoxin gene AaIT1 and the agritol insecticidal crystal protein gene Cyt2B alpha according to a beauveria bassiana preferred codon; guiding the scorpion neurotoxin gene AaIT1 and the agritol insecticidal crystal protein gene Cyt2B alpha in a fungus expression carrier pBarGPE1; converting established recombinant plasmids pBar-Cyt2B alpha-AaIT1 in the bacterial strain of the wild type beauveria bassiana; integrating the recombinant plasmids pBar-Cyt2B alpha-AaIT1 on the bacterial strain of the wild type beauveria bassiana in a non-homologous recombination manner to obtain the bivalent recombination beauveria bassiana. The bivalent recombination beauveria bassiana capable of killing mosquito larvae has the advantages that an effect of killing the mosquito larvae is high.

Description

technical field [0001] The method involves the introduction of bacteria modified by foreign genetic material, specifically the introduction of recombinant Beauveria bassiana with foreign gene modification, and the recombinant Beauveria bassiana can be used to poison and kill mosquito larvae. Background technique [0002] Mosquitoes are important disease vectors, capable of transmitting diseases such as malaria, filariasis, St. Louis encephalitis, Japanese encephalitis, dengue fever, West Nile fever, and yellow fever. Mosquito-borne diseases have a wide range of prevalence, strong transmission power, and high incidence. Every year, hundreds of millions of people around the world are infected with mosquito-borne diseases such as malaria and dengue fever, and the death toll exceeds one million. my country has known 18 genera, 371 species and subspecies of mosquitoes, covering the main vectors of malaria, dengue fever, Japanese encephalitis and filariasis. According to the repo...

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Problems solved by technology

[0007] Interestingly, however, the gpdA promoter and trpC terminator were used to express subtilisin-like protease Pr1 and scorpion toxin AaIT in Beauveria bassiana, but no synergistic virulence effect was found. The reason may be that AaIT is blocked by Pr1 in the blood cavity. Digestion (Lu DD, Pava-Ripoll M, Li ZZ, Wang CS. Insecticidal evaluation of Beauveria bassiana engineered to express a scorpion neurotoxin and a cuticle degrading protease. Appl Microbiol Biotechnol. 2008, 81:515–522)

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