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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: 2016-09-14
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 attack of Lepidoptera pests such as corn borer and cotton bollworm. However, there is no report about controlling the damage of oriental armyworm to plants by producing transgenic plants expressing Cry2Ab protein.

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0085] The first embodiment, the acquisition and synthesis of genes

[0086] 1. Obtain the nucleotide sequence

[0087] 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.

[0088] 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.

[0089] 2. Synthesize the above nucleotide sequence

[0090] 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

[0091] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0092] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0093] 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 Nucleotide sequence (SEQ ID NO: 2); MCS is multiple cloning site).

[0094]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 competent c...

no. 3 example

[0103] The third embodiment, the acquisition of transgenic plants

[0104] 1. Obtaining transgenic corn plants

[0105] 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 DBN100033 and DBN100029 (including CaMV35S promoter sequence, Cry2Ab nucleotide sequence, Cry1A.105 nucleotide sequence, Hpt gene and Nos terminator sequence) were transferred into the maize genome, and obtained The corn plant transformed with Cry2Ab nucleotide sequence and the corn plant transformed with Cry2Ab-Cry1A.105 nucleotide sequence; at the same time, the wild-type corn plant was used as a control.

[0106] For Agrobacterium-mediated maize transformation, briefly, immature immature embryos were isolated from maize and contacted with a suspension of Agrobacterium capable of conve...

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Abstract

The invention relates to an application of insecticidal protein. A method for controlling mythimna seperata pests comprises the following step: the mythimna seperata pests are at least contacted with Cry2Ab protein. According to the method, the mythimna seperata pests are controlled through the Cry2Ab protein which is generated in a plant body and capable of killing mythimna seperata, compared with an agricultural control method, a chemical control method and a physical control method used in the prior art, whole growth period and whole plant protection is performed on the plant so as to control attacks of the mythimna seperata pests, and the method is pollution-free, residual-free, stable and thorough in effect, simple, convenient and economical.

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 oriental armyworm damage to plants. Background technique [0002] Oriental armyworm (Mythimna seperata) belongs to Lepidoptera Noctuidae, is a kind of omnivorous, migratory, intermittent explosive pest that mainly damages wheat, corn, sorghum, soybean, rice and other food crops and pastures. In addition to Xinjiang, which has not yet been reported, there are hazards in other places in my country. Eastern armyworm likes to eat leaves, and after the third instar, it can bite the whole leaf into a notch shape, or eat all the heart leaves to form heartless seedlings. After the third instar, it reaches the overeating period, and can eat up all the aboveground parts of the seedlings, or eat the entire leaves of the plants, leaving only the veins, resulting in severe yield reduction, or even no harvest. In addi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N47/44A01P7/04C12N15/84A01H5/00A01H1/02A01H4/00
Inventor 陶青庞洁杨旭岳健婷张欣馨
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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