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Biological insecticide screening model and its preparing method and application

A biopesticide and model technology, applied in the field of pesticides, can solve the problems of low sensitivity, unknown target and mechanism, and affecting screening efficiency

Inactive Publication Date: 2007-08-15
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, the screening of insect growth regulators usually takes the whole insect body as the object. This screening method has weak screening signal, low sensitivity, and unknown target and mechanism, which greatly affects the screening efficiency.

Method used

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  • Biological insecticide screening model and its preparing method and application
  • Biological insecticide screening model and its preparing method and application
  • Biological insecticide screening model and its preparing method and application

Examples

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preparation example

[0145] 1. Materials:

[0146] 1. EcR sequence (Genbank ID: AF210733), USP sequence (Genbank ID: AF210734) was replaced with the preferred codon of Pichia pastoris, the response element of Drosophila ecdysone receptor was repeated 5 times, 5×EcRE (using Drosophila The EcRE sequence of the HSP27 promoter region, Genbank ID: Y07758), were all synthesized by Jerui Bioengineering (Shanghai) Co., Ltd.;

[0147] 2. Primers: designed with DNAMAN software and synthesized by Beijing Saibaisheng Company.

[0148] Primer 1: 5`-gagctccccacaacaccatatag-3`

[0149] Primer 2: 5`-gaattccgcccttagattag-3`

[0150] Primer 3: 5`-tctagacacgtccgacggc-3`

[0151] Primer 4: 5`-aagcttagcttgcaaattaaag-3`

[0152] 3. pGAPZ plasmid: purchased from Invitrongen

[0153] 4. pPIC3.5K plasmid: purchased from Invitrongen

[0154] 5. pEGFP-N1: purchased from American clonetech company

[0155] 6. Pichia pastoris GS115: purchased from Invitrongen

[0156] 7. The tool enzymes HindIII, Sac I, Bgl II, and No...

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Abstract

The invention discloses a biological insecticide screening mode, which comprises the following steps: integrating acceptor EcR with ecdysone GAP with gene expressing box Promoter-EcR-AOX1 TT and super spiral protein USP gene with gene expressing box TEF1 Promoter-USP-CYC1 TT on the colorant layer of Pasteur pichia; integrating DNA fragment with fruit fly 5XEcRE-HSP27 Promoter-GFP on the colorant layer yeast; getting the product. To co-culture screening drug with this product, we can get stopping or reinforcing GFP expression biological insecticide.

Description

field of invention [0001] The invention relates to the field of pesticides, in particular to a method for large-scale screening of biopesticides using genetic engineering methods. Background technique [0002] With the change of global natural environment and the impact of human activities, the spread of insect-borne diseases tends to increase. At present, the control of hygienic pests mainly uses chemical killing methods. Although chemical insecticides are highly toxic to hygienic pests and aquatic organisms, they are also harmful to mammals, and hygienic pests are prone to develop resistance to chemical insecticides. [0003] Insect growth regulator is a biological insecticide developed in the past 20 years. It is safe for the environment and humans and animals, and has a good control effect. It can also prevent insects from developing resistance and achieve long-term control purposes. Therefore, the application prospect is very broad. However, at present, the screening o...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/12C12R1/84
Inventor 彭鸿娟陈晓光顾金保杨俊
Owner SOUTHERN MEDICAL UNIVERSITY
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