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Application of insecticidal protein

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

Active Publication Date: 2015-01-21
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 (Lepidoptera) pests such as corn borer and cotton bollworm. However, there is no report about controlling the 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

[0091] The first embodiment, the acquisition and synthesis of Cry2Ab gene

[0092] 1. Obtain the Cry2Ab nucleotide sequence

[0093] 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.

[0094] 2. Obtain the nucleotide sequence of Cry1A.105

[0095] 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.

[0096] 3. Synthesize the above nucleotide sequence

[0097] 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

[0098] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0099] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0100] 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 nucleus Nucleotide sequence (SEQ ID NO: 2); MCS is a multiple cloning site).

[0101]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 c...

no. 3 example

[0110] The third embodiment, the acquisition and verification of corn plants transferred to the Cry2Ab gene

[0111] 1. Obtaining corn plants transferred to the Cry2Ab gene

[0112] 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.

[0113] For Agrobacterium-mediated maize transformation, briefly, immat...

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Abstract

The invention relates to an application of insecticidal protein, a method for controlling sesamia inferen insect is characterized in that sesamia inferen insect is contacted to Cry2Ab protein. The method is characterized in that Cry2Ab protein capable of killing sesamia inferen insect is generated in transgenic plants to control the sesamia inferen insect. Compared with an agricultural control method, a chemical control method and a biological control method in prior art, the method can realize whole growth period and whole plant protection on the plants, can prevent and treat sesamia inferen insect, and has the advantages of no pollution, no residues, stable and thoroughly effect, simpleness, convenience and economy.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry2Ab protein expressed in plants to control the plant damage caused by the large borer. Background technique [0002] Sesamia inferens belongs to Lepidoptera Noctuidae. It is an omnivorous pest. In addition to maize, it also damages rice, sugarcane, wheat, sorghum and other gramineous crops. It is widely distributed in central and southeastern my country, especially in Shaanxi. , Most of the rice areas south of Henan. The borer larvae bore into the stems of crops and cause damage to dead core seedlings or the whole plant. The boreholes are generally large, and a large amount of insect feces are discharged out of the stems. Occurs heavier than spring corn. [0003] Corn and sorghum are important food crops in China. The annual loss of food due to the large borer is huge, and even affects the living conditions of the local population. In order to preven...

Claims

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Application Information

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IPC IPC(8): A01N57/16A01N47/44A01P7/04C12N15/84A01H1/02A01H5/00A01H5/10
CPCA01H5/10A01N47/44A01N63/50
Inventor 李建勇杨旭张爱红张欣馨李梅
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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