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Application of insecticidal protein

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

Active Publication Date: 2015-08-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] It has been proved that plants transgenic with Cry1A.105 gene can resist Lepidoptera pests such as corn borer. However, there is no report on the control of sorghum barborer damage to plants by producing transgenic plants expressing Cry1A.105 protein.

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

[0082] First Example, Acquisition and Synthesis of Gene

[0083] 1. Obtain the nucleotide sequence

[0084] The amino acid sequence (1177 amino acids) of the Cry1A.105 insecticidal protein is shown in SEQ ID NO: 1 in the sequence listing; the Cry1A.105 nucleotide sequence corresponding to the amino acid sequence of the Cry1A.105 insecticidal protein ( 3534 nucleotides), as shown in SEQ ID NO: 2 in the sequence listing.

[0085] The amino acid sequence (634 amino acids) of the coding Cry2Ab insecticidal protein, as shown in SEQ ID No: 3 in the sequence listing; the Cry2Ab nucleotide sequence (1905 nucleotides) corresponding to the amino acid sequence of the Cry2Ab insecticidal protein. ), as shown in SEQ ID NO: 4 in the sequence listing.

[0086] 2. Synthesis of the above nucleotide sequence

[0087] The Cry1A.105 nucleotide sequence (as shown in SEQ ID NO: 2 in the sequence listing) and the Cry2Ab nucleotide sequence (as shown in SEQ ID NO: 4 in the sequence listing) were p...

no. 2 example

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

[0089] 1. Construction of recombinant cloning vector containing Cry1A.105 gene

[0090] The synthesized Cry1A.105 nucleotide sequence was ligated into 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 , the construction process is as follows figure 1 (wherein, Amp represents the ampicillin resistance gene; f1 represents the origin of replication of phage f1; LacZ is the LacZ initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; Cry1A.105 is Cry1A .105 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0091] Then the recombinant cloning vector DBN01-T was transformed into E. coli T1 competen...

no. 3 example

[0101] The third embodiment, the acquisition of transgenic plants

[0102] 1. Obtaining transgenic corn plants

[0103] According to the conventional Agrobacterium infection method, the immature embryos of aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second embodiment, so that the embryos constructed in 2 in the second embodiment were co-cultured. T-DNA (including the promoter sequence of maize Ubiquitin gene, Cry1A.105 nucleotide sequence, Cry2Ab nucleotide sequence, Hpt gene and Nos terminator sequence) in the recombinant expression vectors DBN100745, DBN100744 and DBN100029 were transferred into the maize chromosome In the group, corn plants transformed with Cry1A.105 nucleotide sequence, corn plants transformed with Cry2Ab nucleotide sequence, and corn plants transformed with Cry1A.105-Cry2Ab nucleotide sequence were obtained; as comparison.

[0104] For Agrobacterium-mediated transformation of maize, brie...

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Abstract

The invention relates to an application of insecticidal protein. A method for controlling chilo sacchariphagus pests comprises the step as follows: the chilo sacchariphagus pests are at least contacted with Cry1A.105 protein. According to the application of the insecticidal protein, the chilo sacchariphagus pests are controlled by the Cry1A.105 protein which is generated in plant bodies and capable of killing chilo sacchariphagus; compared with an agricultural control method, a chemical control method and a physical control method in the prior art, the method adopted by the insecticidal protein protects the whole plants in the whole growth period so as to control invasion of the chilo sacchariphagus pests, and the insecticidal protein is pollution-free, residue-free, stable and thorough in effect, simple, convenient and economical.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry1A.105 protein to control the damage to plants by Sorghum sorghum by expressing it in a plant. Background technique [0002] The sorghum strip borer Chilo sacchariphagus belongs to the order Lepidoptera, Moth family. Also known as sugar cane borer, or sorghum borer. Mainly distributed in East Asia, South Asia, Southeast Asia and the Indian Ocean. It is widely distributed in China and occurs in most areas of Northeast China, North China, East China and South China. It can harm crops such as corn, sorghum, millet, hemp, and sugarcane. In the dry grain areas of the north, it mainly damages crops such as corn, sorghum and millet, and often occurs in combination with corn borer, resulting in dead heart seedlings; it is also the main pest on sugarcane in the south. [0003] Corn is an important food crop in China. With the strengthening of the global gre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01P7/04C12N15/84A01H5/00A01H1/02A01H4/00A01G13/00A01G1/00
Inventor 张爱红杨旭
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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