Application of insecticidal protein

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

Active Publication Date: 2015-09-23
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 Cry2Ab gene can resist the invasion of Spodoptera frugiperda pest. However, there is no report on the control of sorghum barellar damage to plants by producing transgenic plants expressing Cry2Ab 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

[0083] The first embodiment, the acquisition and synthesis of genes

[0084] 1. Obtain the nucleotide sequence

[0085] The amino acid sequence (634 amino acids) of Cry2Ab insecticidal protein, as shown in SEQ ID NO:1 in the sequence listing; The Cry2Ab nucleotide sequence (1905 nucleotides) of encoding corresponding to the amino acid sequence of described Cry2Ab insecticidal protein , as shown in SEQ ID NO:2 in the sequence listing.

[0086] The amino acid sequence (1177 amino acids) of Cry1A.105 insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Cry1A.105 nucleotide sequence ( 3534 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0087] 2. Synthesize the above nucleotide sequence

[0088] The Cry2Ab nucleotide sequence (as shown in SEQ ID NO: 2 in the sequence listing) and the Cry1A.105 nucleotide sequence (as shown in SEQ ID NO: 4 in the sequence listing) were provided by Nanjing GenScript Co., Ltd.; the 5' end of the synthetic Cry2Ab...

no. 2 example

[0089] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0090] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0091] The synthetic Cry2Ab nucleotide sequence was connected 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, which Build process such as 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; Cry2Ab is the Cry2Ab nucleoside acid sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0092]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 co...

no. 3 example

[0102] The third embodiment, the acquisition of transgenic plants

[0103] 1. Obtaining transgenic corn plants

[0104] 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 The T-DNA in recombinant expression vectors DBN100744, DBN100745 and DBN100029 (including the promoter sequence of maize Ubiquitin gene, Cry2Ab nucleotide sequence, Cry1A.105 nucleotide sequence, Hpt gene and Nos terminator sequence) were transferred to maize chromosome In the group, the corn plants transferred to Cry2Ab nucleotide sequence, the corn plant transferred to Cry1A.105 nucleotide sequence and the corn plant transferred to Cry1A.105-Cry2Ab nucleotide sequence were obtained; as comparison.

[0105] For Agrobacterium-mediated transformation of maize, briefly, immature immature embryos were isolated from maize and contact...

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Abstract

The invention relates to application of an insecticidal protein. A method for controlling sorghum striped borer pests comprises the step of at least contacting the sorghum striped borer pests with a Cry2Ab protein. The sorghum striped borer pests are controlled by generating the Cry2Ab protein in a plant body to kill the sorghum striped borer pests. Compared with the agricultural prevention and control method, chemical prevention and control method and physical prevention and control method in the prior art, the method has the advantages that the whole plant is protected in the whole growth period to prevent and control the invasion of the sorghum striped borer pests, pollution and residue are not caused, the effect is stable and thorough, and the method 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 Cry2Ab protein expressed in plants to control damage to plants by the sorghum borer. Background technique [0002] The sorghum bar borer Chilo sacchariphagus belongs to the order Lepidoptera and belongs to the Mothidae family. Also known as sugarcane borer, or sorghum borer. Mainly distributed in East Asia, South Asia, Southeast Asia and the Indian Ocean region. The domestic distribution is very wide, and it occurs in most parts of Northeast, North China, East China and South China. Can harm corn, sorghum, millet, hemp, sugar cane and other crops. In dry grain areas in the north, it mainly damages crops such as corn, sorghum and millet, and often occurs in combination with corn borer, causing dead 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01P7/04A01H5/00A01H1/02A01H4/00A01G13/00A01G1/00C12N15/82C12N15/84C12N15/89C12N15/87A01N63/50
CPCA01N37/18A01H4/00A01N47/44C12N15/80A01G2/00A01N63/50A01H1/127A01N63/23
Inventor 杨旭张爱红
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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