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Construct for controlling insect pests and method thereof

A technology for constructs and pests, applied in the directions of botanical equipment and methods, pesticides, introduction of foreign genetic material using vectors, etc., can solve the problems of increasing production costs, hindering the promotion of genetically modified resistant materials, and requiring repeated application, etc.

Active Publication Date: 2014-04-16
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and need to be repeatedly applied in production, which greatly increases the production cost
[0004] At present, most of the Cry1Ca protein and Cry1Ab / Ac protein are used in plants in the form of a single gene, especially the Cry1Ca protein. When it exists in a single gene form, it has low resistance to Chilorrhizae and S. The emergence of resistance to these two genes alone by the borer and the large borer hampers the promotion of transgenic resistant materials

Method used

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  • Construct for controlling insect pests and method thereof
  • Construct for controlling insect pests and method thereof
  • Construct for controlling insect pests and method thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0082] The first embodiment, the acquisition and synthesis of Cry1Ca gene and Cry1Ab / Ac gene

[0083] 1. Obtain the nucleotide sequences of Cry1Ca and Cry1Ab / Ac

[0084] The amino acid sequence (630 amino acids) of the Cry1Ca insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; the Cry1Ca-01 nucleotide sequence encoding the amino acid sequence (630 amino acids) corresponding to the Cry1Ca insecticidal protein (1896 nucleotides), as shown in SEQ ID NO:3 in the sequence listing, and the nucleotide sequence of Cry1Ca-02 (1896 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0085] The amino acid sequence (609 amino acids) of the Cry1Ab / Ac insecticidal protein, as shown in SEQ ID NO: 2 in the sequence listing; Cry1Ab / Ac encoding the amino acid sequence (609 amino acids) corresponding to the Cry1Ab / Ac insecticidal protein Nucleotide sequence (1830 nucleotides), as shown in SEQ ID NO:5 in the sequence listing.

[0086] 2. Synthesize the above nuc...

no. 2 example

[0088] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0089] 1. Construction of a recombinant cloning vector containing the Cry1Ca gene and the Cry1Ab / Ac gene

[0090] The synthesized Cry1Ca-01 nucleotide sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T , its construction process is as follows figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; Cry1Ca-01 is the promoter of Cry1Ca -01 nucleotide sequence (SEQ ID NO: 3); MCS is a multiple cloning site).

[0091] Then, the recombinant cloning vector...

no. 3 example

[0103] The third embodiment, the acquisition and verification of corn plants transferred to Cry1Ca-01 and Cry1Ab / Ac genes

[0104] 1. Obtain corn plants transferred to Cry1Ca-01 and Cry1Ab / Ac genes

[0105]According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second example, so that the T-DNA in recombinant expression vectors DBN100323, DBN100060, DBN100056 and DBN100062 (including the promoter sequence of corn Ubiquitin gene, Cry1Ca-01 nucleotide sequence, Cry1Ca-02 nucleotide sequence, Cry1Ab / Ac nucleotide sequence, PMI gene and Nos terminator sequence) were transferred into the maize genome, and the maize plants transferred to the Cry1Ca-01 nucleotide sequence, the maize plants transferred to the Cry1Ca-02 nucleotide sequence, and the Cry1Ab / Ac nucleoside The maize plant of the acid sequence and the maize plant of the Cry1Ca-01...

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Abstract

The invention relates to a construct for controlling insect pests and a method thereof. The construct comprises a Cry1Ca protein and a Cry1A protein. According to the construct for controlling insect pests and the method thereof, the protein Cry1Ca and Cry1Ab / Ac are generated in plants so as to control the insect pests. Compared with agricultural control methods, chemical control methods and biological control methods are used in the prior art, the method is high in toxicity, complete in effect, pollution-free, residue-free, stable in effect, simple, convenient and economic, and the plants are subjected to whole-growth period and whole-plant protection so as to prevent intrusion of the insect pests.

Description

technical field [0001] The present invention relates to a construct and method for controlling pests, in particular to a method for controlling pests to damage plants by using a construct expressing Cry1Ca protein and Cry1Ab / Ac protein in plants. Background technique [0002] Corn and rice are important food crops in China. Every year, plant pests cause huge food losses, and even affect the living conditions of the local population. In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and require repeated application in production, which greatly increases production costs. [0003] In order to solve the limitations in the practical application of chemical insecticides and biological ins...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12P21/00A01H5/00A01N47/44A01P7/04
Inventor 韩超庞洁丁德荣李胜兵岳健婷
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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