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Use of insecticidal protein

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

Active Publication Date: 2016-06-15
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Plants transgenic for Vip3A have been shown to be resistant to Lepidoptera pests such as cutworm, cotton bollworm, and fall armyworm. Hazard Reporting

Method used

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  • Use of insecticidal protein
  • Use of insecticidal protein
  • Use of insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0084] The first embodiment, the acquisition and synthesis of genes

[0085] 1. Obtain the nucleotide sequence

[0086] The amino acid sequence (789 amino acids) of Vip3A-01 insecticidal protein, as shown in SEQIDNO: 1 in the sequence listing; The Vip3A-01 nucleotide sequence (2370 of coding corresponding to the aminoacid sequence of described Vip3A-01 insecticidal protein Nucleotide), as shown in SEQIDNO:2 in the sequence listing.

[0087] The amino acid sequence (789 amino acids) of Vip3A-02 insecticidal protein, as shown in SEQIDNO:3 in the sequence listing; The Vip3A-02 nucleotide sequence (2370 of coding corresponding to the aminoacid sequence of described Vip3A-02 insecticidal protein Nucleotide), as shown in SEQ ID NO:4 in the sequence listing.

[0088] The amino acid sequence (1436 amino acids) of Vip3A+Cry1Ab insecticidal protein, as shown in SEQIDNO: 9 in the sequence table; The Vip3A+Cry1Ab nucleotide sequence (4311) of encoding corresponding to the amino acid seq...

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 a multiple cloning site).

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

no. 3 example

[0108] The third embodiment, the acquisition of transgenic plants

[0109] 1. Obtaining transgenic corn plants

[0110] 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, DBN100740, DBN100736 and DBN100003 (including the promoter sequence of corn Ubiquitin gene, Vip3A-01 nucleotide sequence, Vip3A-02 nucleotide sequence, Vip3A+Cry1Ab nucleotide sequence, Cry1Ab- 01 nucleotide sequence, Hpt gene and Nos terminator sequence) were transferred into the maize genome, and the maize plant transferred to the Vip3A-01 nucleotide sequence, the maize plant transferred to the Vip3A-02 nucleotide sequence, Maize plants transfected with Vip3A+Cry1Ab nucleotide sequence, maize plants transfected with Vip3A-01-Cry1Ab-01 nucleotide sequence; meanwhile, wild-type ma...

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Abstract

The invention relates to a use of an insecticidal protein; a method for controlling conogethes punctiferalis pests comprises the steps: allowing the conogethes punctiferalis pests at least to make contact with a Vip3A protein. The Vip3A protein which can kill the conogethes punctiferalis is generated in plants to control the conogethes punctiferalis pests; compared with an agriculture prevention and treatment method, a chemical prevention and treatment method and a physical prevention and treatment method used in the prior art, the method performs whole growth period and whole plant protection of the plants to prevent and treat infringement of the conogethes punctiferalis pests, has the advantages of no pollution, no residue, and stable and thorough effect, and 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 peach borer borer. Background technique [0002] The peach borer Conogethes punctiferalis belongs to Lepidoptera Mothidae. It is an omnivorous pest. It not only harms corn, sorghum and other crops, but also harms peaches, persimmons, chestnuts and other fruit trees. It is widely distributed in my country, from Heilongjiang and Inner Mongolia in the north to The southern edge of Taiwan, Hainan, Guangdong, Guangxi, and Yunnan, the east of the former Soviet Union, and the north of Korea, and the west from Shanxi and Shaanxi to Ningxia and Gansu, and turns into Sichuan, Yunnan, and Tibet. When maize is harmed, it mainly eats ears and stems, and the plant damage rate reaches 30%-80%. When sorghum is harmed, newly hatched larvae eat into young sorghum grains and seal their mouths with fec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01N57/16A01P7/04A01G13/00
CPCA01G13/00A01N47/44A01N57/16
Inventor 杨旭张爱红
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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