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Application of insecticidal protein

A protein and pest technology, applied in the field of insecticidal protein

Active Publication Date: 2020-06-19
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Plants transgenic for the Vip3Aa gene have been shown to be resistant to pests of the order Lepidoptera such as cutworms, cotton bollworms, and fall armyworms, however, so far there is no research on the control of South American bollworms by producing transgenic plants expressing the Vip3Aa protein. Reports of damage to plants

Method used

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  • Application of insecticidal protein
  • Application of insecticidal protein
  • Application of insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0085] The first embodiment, gene acquisition and synthesis

[0086] 1. Obtain the nucleotide sequence

[0087] The amino acid sequence (789 amino acids) of the Vip3Aa-01 insecticidal protein is shown in SEQ ID NO:1 in the sequence table; the nucleotide sequence of Vip3Aa-01 encoding the amino acid sequence of the Vip3Aa-01 insecticidal protein ( 2370 nucleotides), as shown in SEQ ID NO: 2 in the sequence listing.

[0088] The amino acid sequence (789 amino acids) of the Vip3Aa-02 insecticidal protein is shown in SEQ ID NO: 3 in the sequence table; the Vip3Aa-02 nucleotide sequence encoding the amino acid sequence of the Vip3Aa-02 insecticidal protein ( 2370 nucleotides), as shown in SEQ ID NO: 4 in the sequence listing.

[0089] The amino acid sequence (789 amino acids) of the Vip3Aa-03 insecticidal protein is shown in SEQ ID NO: 5 in the sequence table; the nucleotide sequence of Vip3Aa-03 encoding the amino acid sequence of the Vip3Aa-03 insecticidal protein ( 2370 nucleotides), ...

no. 2 example

[0095] The second embodiment, construction of recombinant expression vector and transformation of Agrobacterium with recombinant expression vector

[0096] 1. Construct a recombinant cloning vector containing Vip3Aa gene

[0097] Connect the synthesized Vip3Aa-01 nucleotide sequence to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and proceed according to the instructions of Promega product pGEM-T vector to obtain the inverted recombinant cloning vector DBN01- T, its construction process is as figure 1 As shown (where Amp represents the ampicillin resistance gene; flori represents the origin of replication of phage fl; LacZ is the actual codon of LacZ; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; Vip3Aa-01 is Vip3Aa-01 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0098] Then the recombinant cloning vector DBN01-T was transformed into E. coli T1 competent cells (Transgen, Beijing, China, CAT: CD501) by the ...

no. 3 example

[0115] The third embodiment, the acquisition of transgenic plants

[0116] 1. Obtain genetically modified soybean plants

[0117] According to the conventional Agrobacterium infection method, the cotyledonary node tissue of the aseptically cultured soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 3 in the second embodiment to replace the recombinant T-DNA expression vectors DBN100002, DBN100741, DBN100742, DBN100743, DBN100003 and DBN100370 (including the promoter sequence of the Arabidopsis ubiquitin gene, the nucleotide sequence of Vip3Aa-01, the nucleotide sequence of Vip3Aa-02, the nucleotide sequence of Vip3Aa-03 Nucleotide sequence, Vip3Aa-04 nucleotide sequence, Vip3Aa-02-Cry1Ab nucleotide sequence, Vip3Aa-01-Cry2Ab nucleotide sequence, PAT gene and tNos terminator sequence) were transferred into the soybean genome, and obtained Soybean plants transformed with the nucleotide sequence of Vip3Aa-01, soybean plants transformed with the nucleoti...

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Abstract

The application of the insecticidal protein comprises the following steps: enabling cotton bollworm pests of South America to be at least in contact with Vip3Aa protein, and controlling the cotton bollworm pests of South America by generating the Vip3Aa protein capable of killing the cotton bollworms in plants.

Description

Technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Vip3Aa protein to control South American cotton bollworm damage to plants by expressing it in plants. Background technique [0002] The South American cotton bollworm Helicoverpa gelotopoeon belongs to the genus Bollworm of the Lepidoptera Noctuidae. It is mainly distributed in southern South America such as Brazil, Argentina, Bolivia, Paraguay and Uruguay. Helicoverpa armigera is a polyphagous pest. It mainly damages cash crops such as soybean, cotton, firstfa, sunflower, chickpea and corn. Its characteristics are: the whole larval stage can damage most plants such as stalks, leaves, inflorescences and fruits organization. Like eating soybeans, its larvae can feed on 81.8% of the aboveground tissues of soybeans, and consume 340cm in the larval stage 2 Soybean leaves, the last larvae can consume 15 soybean seeds, which will seriously affect the yield. [0003]...

Claims

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Application Information

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IPC IPC(8): A01N37/46A01P7/04A01G13/00
CPCA01N37/46A01G13/00
Inventor 韩超于彩虹谢香庭周毅刘海利
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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