Engineering strain producing anti-fungal protein genes and application

A technology of genetically engineered strains and antibacterial proteins, applied in the field of genetic engineering, can solve the problems of poor stability and low yield, and achieve the effects of short time, high recovery rate and high expression rate

Inactive Publication Date: 2016-01-20
INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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  • Engineering strain producing anti-fungal protein genes and application
  • Engineering strain producing anti-fungal protein genes and application
  • Engineering strain producing anti-fungal protein genes and application

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Embodiment Construction

[0019] The technical solution of the present invention will be further described below in conjunction with the accompanying drawings, but it is not limited thereto. Any modification or equivalent replacement of the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention should be covered by the present invention. within the scope of protection.

[0020] The invention provides a genetically engineered bacterial strain producing antimicrobial protein, comprising the following contents:

[0021] 1. Antibacterial protein gene synthesis and construction of genetically engineered strains

[0022] According to the genome sequence of the strain, the antibacterial protein gene was artificially synthesized, and the gene 5 ， Add a BamHI restriction site at the end, at 3 ， The XholI restriction site is added to the end, and the gene sequence of the synthesized antibacterial protein AP1 is shown in SEQ ID ...

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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. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12P21/02A01N63/00A01P3/00C12R1/19
Inventor 姜威张淑梅李晶孟立强刘宇帅胡基华陈静宇曹旭
Owner INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
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