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Application of insecticidal protein

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

Active Publication Date: 2015-01-07
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] It has been proved that the plants transgenic for Cry2Ab can resist the damage of Lepidoptera pests such as corn borer and cotton bollworm. to report

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 Cry2Ab gene

[0091] 1. Obtain the Cry2Ab nucleotide sequence

[0092] 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.

[0093] 2. Obtain the nucleotide sequence of Cry1A.105

[0094] The Cry1A.105 nucleotide sequence (3534 nucleotides) encoding the amino acid sequence (1177 amino acids) of the Cry1A.105 insecticidal protein is shown in SEQ ID NO:3 in the sequence listing.

[0095] 3. Synthesize the above nucleotide sequence

[0096] 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: 3 in the sequence listing) were provided by Nanjing GenScript Biotechnolog...

no. 2 example

[0097] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0098] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0099] 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).

[0100]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 co...

no. 3 example

[0109] The third embodiment, the acquisition and verification of corn plants transferred to the Cry2Ab gene

[0110] 1. Obtaining corn plants transferred to the Cry2Ab gene

[0111] 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 the recombinant expression vectors DBN100033 and DBN100076 (including the promoter sequence of the maize Ubiquitin gene, the Cry2Ab nucleotide sequence, the Cry1A.105 nucleotide sequence, the PMI gene and the Nos terminator sequence) were transferred into the maize genome , obtained the corn plant transformed into the Cry2Ab nucleotide sequence and the corn plant transformed into the Cry2Ab-Cry1A.105 nucleotide sequence; at the same time, the wild type corn plant was used as a control.

[0112] For Agrobacterium-mediated maize transformation, briefly, immat...

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Abstract

The invention relates to an application of an insecticidal protein. A method for controlling athetis lepigone comprises a step of contacting athetis lepigone with a Cry2Ab protein. The method adopts a transgene technology to generate Cry2Ab proteins, which can kill the athetis lepigone, in plants to achieve the goal of controlling athetis lepigone. Compared to the conventional agricultural control method and chemical control method, the provided method can protect whole plants from athetis lepigone during the whole growth period, and moreover has the advantages of no pollution, no residue, stable and radical effect, simpleness, convenience, and economy.

Description

technical field [0001] The invention relates to an application of an insecticidal protein, in particular to an application of a Cry2Ab protein to control plant damage to plants by expressing Cry2Ab protein in plants. Background technique [0002] Athetis lepigone belongs to Lepidoptera Noctuidae and is an omnivorous pest. At present, it is found that it mainly harms corn crops. It is mainly distributed in the summer corn planting areas of my country's Huanghuaihai, and abroad mainly in Japan, North Korea, Russia, Europe and other places. The two-spot moth mainly harms the corn roots at the soil surface of the aerial roots of corn, and bites off the corn stems or superficial roots of the corn. A large area of ​​blank land appears, seriously endangering plots and even destroying species. [0003] Corn is the main food crop in China. On July 9, 2011, CCTV News Network reported for the first time the occurrence of Noctuid moth in China. From the autumn of 2011 to May 31, 2012, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01N57/16A01P7/04C12N15/84A01H5/00A01H1/02A01H5/10
Inventor 丁德荣陶青张欣馨杨旭康越景
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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