Engineering strain producing anti-fungal protein genes and application

Abstract

The invention discloses an engineering strain producing anti-fungal protein genes and a construction method and application thereof. The strain is pET32a(+)-Ap1-BL21, and the preservation serial number is CGMCC No.11542. According to the construction method, double enzyme digestion is carried out on anti-fungal protein AP1 genes and plasmid pET32a(+) through BamH I and XholI, and the anti-fungal protein AP1 genes and the plasmid pET32a(+) are connected through T4DNA ligase and then converted into E.coli BL21 competent cells to obtain the engineering strain pET32a(+)-Ap1-BL21 with the anti-fungal protein genes. The gene engineering strain contains the anti-fungal protein genes, induction time at the temperature of 30 DEG C is short, the expression rate (43.2%) is high, the purification protein yield (124.82 mg / L) is high, the recovery rate (92%) is high, protein has a good inhibiting effect on phytopathogen such as soybean root rot bacteria and fusarium fujikuroi after ITPG inducting, HIS column purifying and enterokinase restriction enzyme digesting are carried out, and the strain provides a novel strain resource for research and development of recombinant protein pesticide.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a gene for producing an antibacterial protein, an engineering strain and its application. Background technique [0002] With the continuous development of sustainable agriculture and the increasingly serious problems of chemical pesticide resistance and pollution, microbial pesticides have become an important means of plant disease control. At present, the widely used microbial pesticides are mainly live bacterial preparations, which have short storage periods and unstable control effects. Studies have found that some biocontrol bacteria can produce antibacterial proteins, but it is difficult to industrialize due to low yield and poor stability. In recent years, with the continuous development of molecular biology and biotechnology, the research and development of microbial pesticides has developed from live bacterial preparations to recombinant microbial pesticides. ...

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Problems solved by technology

Studies have found that some biocontrol bacteria can produce antibacterial proteins, but it is difficult to industrialize due to low yield and poor stability

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