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The use of insecticidal protein

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

Active Publication Date: 2018-06-19
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 plants transgenic for Cry1A can resist Lepidoptera pests such as corn borer, cotton bollworm, and fall armyworm. to report

Method used

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  • The use of insecticidal protein
  • The use of insecticidal protein
  • The use of insecticidal protein

Examples

Experimental program
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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 (818 amino acids) of Cry1Ab-01 insecticidal protein, as shown in SEQ ID NO:1 in the sequence listing; Cry1Ab-01 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 nucleotide sequence ( 1848 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0087] The amino acid sequence (1156 amino acids) of Cry1Ac-01 insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1Ac-01 nucleotide sequence ( 3471 nucleotides), as shown in SEQ ID NO:6 in the sequence listing. The amino acid sequence (616 amino acids) of Cry1Ac-02 insecticidal protein, as shown in SEQ ID NO:7 in the sequence listing; Cry1Ac-02 nucleotide sequence ( 1851 nucleotides), a...

no. 2 example

[0093] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0094] 1. Construction of a recombinant cloning vector containing the Cry1A gene

[0095] 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).

[0096] Then, the recombinant cloning vector DBN01-T was transformed into Escher...

no. 3 example

[0115] The third embodiment, the acquisition of transgenic plants

[0116] 1. Obtaining transgenic corn plants

[0117] 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 in recombinant expression vectors DBN100124, DBN100743, DBN100741, DBN100734, DBN100737, DBN100735 and DBN100013 (including the promoter sequence of corn Ubiquitin gene, Cry1Ab-01 nucleotide sequence, Cry1Ab-02 nucleotide sequence, Cry1Ac-01 Nucleotide sequence, Cry1Ac-02 nucleotide sequence, Cry1Ie nucleotide sequence, Cry1Ab / Ac nucleotide sequence, Cry1Fa nucleotide sequence, Hpt gene and Nos terminator sequence) are transferred into the maize genome to obtain Corn plants transferred into Cry1Ab-01 nucleotide sequence, corn plants transferred into Cry1Ab-02 nucleotide sequence, corn plants transferred into Cry1Ac-01 nucleo...

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Abstract

The invention relates to an application of an insecticidal protein, and the method for controlling the pest of the sorghum borer comprises: at least contacting the pest of the sorghum borer with the Cry1A protein. The present invention controls the sorghum borer pest by producing the Cry1A protein capable of killing the sorghum borer in the plant; compared with the agricultural control methods, chemical control methods and physical control methods used in the prior art, the present invention controls the plants during the whole growth period , The protection of the whole plant is used to prevent and control the damage of the sorghum barborer pest, and it has no pollution and no residue, the effect is stable, thorough, 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 protein to control the damage of plants by the sorghum stalk borer through expression in plants. 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N47/44A01P7/04C12N15/84A01H5/00A01H1/02A01H4/00A01G13/00A01G22/55A01G22/20
CPCA01G13/00A01H4/00A01N47/44A01G2/00
Inventor 杨旭张爱红
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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