Application of insecticidal protein

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

Active Publication Date: 2015-04-08
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] It has been proved that the plants transfected with Vip3A gene can resist the invasion of Lepidoptera pests such as cutworm, fall armyworm, large borer, and corn armyworm. Reports on Controlling Plant Hazards 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

[0086] The first embodiment, the acquisition and synthesis of genes

[0087] 1. Obtain the nucleotide sequence

[0088] Amino acid sequence (789 amino acids) of Vip3A-01 insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; coding corresponds to the Vip3A-01 of the amino acid sequence (789 amino acids) of said Vip3A-01 insecticidal protein Nucleotide sequence (2370 nucleotides), as shown in SEQ ID NO:2 in the sequence listing. Amino acid sequence (789 amino acids) of Vip3A-02 insecticidal protein, as shown in SEQ ID NO: 3 in the sequence listing; coding corresponds to the Vip3A-02 of the amino acid sequence (789 amino acids) of said Vip3A-02 insecticidal protein Nucleotide sequence (2370 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0089] The amino acid sequence (615 amino acids) of Cry1Ab insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; The Cry1Ab nucleotide sequence (1848 nucleotides), as shown in SEQ ID NO:6 in ...

no. 2 example

[0092] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0093] 1. Construction of a recombinant cloning vector containing the Vip3A gene

[0094] The synthetic Vip3A-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 initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; Vip3A-01 is the Vip3A -01 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0095]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia...

no. 3 example

[0106] The third embodiment, the acquisition of transgenic plants

[0107] 1. Obtaining transgenic corn plants

[0108] According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured corn variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second embodiment, so that the T-DNA of recombinant expression vectors DBN100002, DBN100003 and DBN100740 (including the promoter sequence of corn Ubiquitin gene, Vip3A-01 nucleotide sequence, Vip3A-02 nucleotide sequence, Cry1Ab nucleotide sequence, Hpt gene and Nos terminator Sequence) was transferred into the corn genome, and the corn plant transferred to the Vip3A-01 nucleotide sequence, the corn plant transferred to the Vip3A-02 nucleotide sequence, and the corn plant transferred to the Cry1Ab+Vip3A-01 nucleotide sequence were obtained ; At the same time, wild-type maize plants were used as controls.

[0109] For Agrobacterium-mediated transformation of m...

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Abstract

The invention relates to application of insecticidal protein. A method for controlling chilotraea infuscatellus insects comprises the step of enabling the chilotraea infuscatellus insects to be at least in contact with Vip3A protein. The application has the characteristic that the chilotraea infuscatellus insects are controlled by generating the Vip3A protein capable of killing chilotraea infuscatellus in a plant body; compared with an agricultural control method, a chemical control method and a physical control method in the prior art, the application has the advantages that whole-growing-period and whole-plant protection is carried out on the plant so as to control the invasion of the chilotraea infuscatellus insects, no pollution is caused, no residue is generated, the effect is stable and thorough, and the application is simple, convenient and economic.

Description

technical field [0001] The invention relates to an application of an insecticidal protein, in particular to an application of a Vip3A protein expressed in plants to control damage to plants by the pod borer. Background technique [0002] Chestnut gray borer (Chilo infuscatellus), also known as sugarcane two-spot borer, belongs to Lepidoptera moth family. It is widely distributed in Southeast Asia, South Asia and China. In China, it mainly damages gramineous crops, such as corn, sorghum, millet and sugarcane. Millet ash borer is a stem-boring pest, which can cause dead plants after the plant is damaged. Under normal circumstances, the spring valley area and the spring-summer valley mixed-seeding area have heavy occurrences, and the summer valley area has light damage. [0003] Corn is an important food crop in China. With the strengthening of the global greenhouse effect, the temperature has continued to rise in the past two years, and the types and numbers of insect pests h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01P7/04A01G13/00A01M99/00A01H1/02A01H4/00A01H5/00C12N15/84
CPCA01N47/44A01N63/50
Inventor 张爱红陶青李建勇王利君
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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