Transgenosis biocontrol engineering fungus for expressing stomach toxicity insecticidal protein by using endogenous security label and construction method of transgenosis biocontrol engineering fungus

A safety labeling, transgenic technology, applied in microorganism-based methods, biochemical equipment and methods, biocides, etc., can solve problems such as inability to grow normally, loss of function, blocked uracil synthesis, etc.

Inactive Publication Date: 2016-07-13
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

This is because ura3 can regulate the synthesis of uracil in cells, and the loss of its function hinders the synthesis of uracil, so it cannot grow normally

Method used

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  • Transgenosis biocontrol engineering fungus for expressing stomach toxicity insecticidal protein by using endogenous security label and construction method of transgenosis biocontrol engineering fungus
  • Transgenosis biocontrol engineering fungus for expressing stomach toxicity insecticidal protein by using endogenous security label and construction method of transgenosis biocontrol engineering fungus
  • Transgenosis biocontrol engineering fungus for expressing stomach toxicity insecticidal protein by using endogenous security label and construction method of transgenosis biocontrol engineering fungus

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Embodiment Construction

[0021] The recombinant engineering strain obtained by the method of the present invention will be described in detail in combination with experiments below.

[0022] The wild strain Bb2860 of Beauveria bassiana, the ura3 knockout strain △ura3, and the engineering strains BbV28 and BbHV8 expressing Vip3Aa1 are all strains preserved or constructed in our laboratory. The construction of BbV28 used the common fungal promoter, PgpdA of Aspergillus nidulans, to drive the expression of vip3Aa1 in Bb2860, and the construction of BbHV8 used the optimized endogenous hydrophobin gene (hyd1) promoter Phydl-tl to drive the high expression of vip3Aa1 in Bb2860 , both screening markers are glufosinate resistance gene bar. Plasmids: pAN52-Phydl-tl-Vip3Aa1 and p0380-ura3, both of which were constructed in the previous research of our laboratory. The ELISA reagent TMBcomponent was purchased from Amresco (Solon, OH, USA), and the rabbit serum polyclonal antibody to Vip3Aa1 protein was provided ...

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Abstract

The invention discloses a transgenosis biocontrol engineering fungus for expressing stomach toxicity insecticidal protein by using an endogenous security label and a construction method of the transgenosis biocontrol engineering fungus.Encoding genes of the insecticidal protein Vip3Aa1 are placed under an endogenous strong promoter, and an expression vector with the insecticidal protein genes is constructed.Beauveria bassiana endogenesis ura3 auxotroph is taken as a security label, and competence blastospore, which carries the expression vector, of the beauveria bassiana endogenesis ura3 auxotroph is shifted into a beauveria bassiana wild strain to obtain a recombinant engineering strain of high-expression insecticidal protein Vip3Aa1.The transgenosis biocontrol engineering fungus for expressing the stomach toxicity insecticidal protein Vip3Aa1 by using the non-exogenous resistance label is BbHUV5, and has stomach toxicity insecticidal activity which does not exist in the beauveria bassiana inherently, moreover, the integral insecticidal effect is greatly improved, and environmental risks of any exogenous herbicide resistance gene label do not exist.The engineering strain construction idea and an engineering strain thereof open up a new technological path for a transgenosis fungus insecticide with development synergy and high environment consistency degree.

Description

technical field [0001] The invention belongs to the field of molecular biology technology, and relates to DNA recombination technology and application in the field of bioengineering technology, in particular to a transgenic biocontrol engineering fungus expressing stomach poison insecticidal protein with endogenous safety markers and a construction method thereof. Background technique [0002] Transgenic engineering technology has been widely used in the genetic improvement of bio-control fungi, and it is the trump card to improve the potential of fungal bio-control. One is through the research and analysis of the functions of anti-stress-related genes, purposely high-express endogenous stress-resistant genes or introduce exogenous stress-resistant genes to improve the stress resistance of strains, such as anti-oxidation, hyperosmotic resistance, high temperature resistance, and UV radiation, resistance to fungicides, resistance to herbicides. The second is to introduce end...

Claims

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Application Information

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IPC IPC(8): C12N1/15C12N15/80A01N63/04A01P7/04C12R1/645
CPCA01N63/30C07K14/325C12N9/88C12N15/80C12Y401/01023
Inventor 冯明光史汉强童森淼
Owner ZHEJIANG UNIV
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