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Application of insecticidal protein

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

Active Publication Date: 2015-04-22
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 transgenic for Cry1A can resist the attack of Lepidoptera (Lepidoptera) pests such as corn borer, cotton bollworm, and fall armyworm. Hazard Reporting

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

[0090] The first embodiment, the acquisition and synthesis of genes

[0091] 1. Obtain Cry and Vip nucleotide sequences

[0092] The amino acid sequence (818 amino acids) of Cry1Ab-01 insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; Cry1Ab-01 coding corresponding to the amino acid sequence (818 amino acids) of said Cry1Ab-01 insecticidal protein Nucleotide sequence (2457 nucleotides), as shown in SEQ ID NO:2 in the sequence listing. The amino acid sequence (615 amino acids) of Cry1Ab-02 insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Cry1Ab-02 corresponding to the amino acid sequence (615 amino acids) of said Cry1Ab-02 insecticidal protein Nucleotide sequence (1848 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0093] The amino acid sequence (1156 amino acids) of the Cry1Ac-01 insecticidal protein, as shown in SEQ ID NO: 5 in the sequence listing; Cry1Ac-01 encoding the amino acid sequence (1156 amino acids) cor...

no. 2 example

[0099] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0100] 1. Construction of a recombinant cloning vector containing the Cry1A gene

[0101] The synthesized Cry1Ab-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; Cry1Ab-01 is the Cry1Ab -01 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0102] Then, the recombinant cloning vector DBN01-T was transformed into Esche...

no. 3 example

[0120] The third embodiment, the acquisition of transgenic plants

[0121] 1. Obtaining transgenic corn plants

[0122] 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 DBN100124, DBN100743, DBN100741, DBN100734, DBN100737 and DBN100736 (including the promoter sequence of corn Ubiquitin gene, Cry1Ab-01 nucleotide sequence, Cry1Ab-02 nucleotide sequence, Cry1Ac-01 nucleotide sequence sequence, Cry1Ac-02 nucleotide sequence, Cry1Ie nucleotide sequence, Cry1Ab / Ac nucleotide sequence, Cry1Ab+Vip3A nucleotide sequence, Hpt gene and Nos terminator sequence) into the maize genome, obtained Corn plants transferred to Cry1Ab-01 nucleotide sequence, corn plants transferred to Cry1Ab-02 nucleotide sequence, corn plants transferred to Cry1Ac-01 nucleotide sequence, tra...

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Abstract

The invention relates to an application of an insecticidal protein. A control method for chilotraea infuscatellus pests comprises the following step of at least contacting the chilotraea infuscatellus pests with a protein CrylA. The chilotraea infuscatellus pests are controlled through generating the protein CrylA capable of killing chilotraea infuscatellus in a plant body. Compared with an agricultural control method, a chemical control method and a physical control method adopted in the prior art, the control method can be used for avoiding invasion of the chilotraea infuscatellus pests by protecting the whole plant in the whole growth period and is pollution-free, residue-free, stable and thorough in effect, simple, convenient and economic.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry1A protein to control the plant damage caused by the pod borer by expressing it in plants. 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...

Claims

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Application Information

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IPC IPC(8): A01N63/02A01N65/44A01P7/04C12N15/84A01H1/02A01H4/00A01G1/00A01G13/00A01N63/50
CPCA01G13/00A01H4/00A01N65/44A01N63/50A01N63/23
Inventor 杨旭丁德荣张爱红陶青李建勇李梅张云珠
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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