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Application of insecticidal protein

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

Active Publication Date: 2015-04-22
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 with Cry1A.105 gene can resist Lepidoptera (Lepidoptera) pests such as corn borer. However, there is no report about controlling the damage to plants by producing transgenic plants expressing Cry1A.105 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

[0086] The first embodiment, the acquisition and synthesis of genes

[0087] 1. Obtain the nucleotide sequence

[0088] The amino acid sequence (1177 amino acids) of the Cry1A.105 insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; Cry1A.105 encoding the amino acid sequence (1177 amino acids) corresponding to the Cry1A.105 insecticidal protein Nucleotide sequence (3534 nucleotides), as shown in SEQ ID NO:2 in the sequence listing.

[0089] The amino acid sequence (634 amino acids) of encoding Cry2Ab insecticidal protein, as shown in SEQ ID No:3 in the sequence listing; The Cry2Ab nucleotide sequence ( 1905 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0090] 2. Synthesize the above nucleotide sequence

[0091] The Cry1A.105 nucleotide sequence (as shown in SEQ ID NO: 2 in the sequence listing) and the Cry2Ab nucleotide sequence (as shown in SEQ ID NO: 4 in the sequence listing) were provided by Nanjing GenScript Co., Ltd. synthesis; t...

no. 2 example

[0092] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0093] 1. Construction of a recombinant cloning vector containing the Cry1A.105 gene

[0094] The synthesized Cry1A.105 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 start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; Cry1A.105 is the promoter of Cry1A .105 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0095]Then, the recombinant cloning vector DBN01-T was transf...

no. 3 example

[0105] The third embodiment, the acquisition of transgenic plants

[0106] 1. Obtaining transgenic corn plants

[0107] 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 DBN100745, DBN100744 and DBN100029 (including the promoter sequence of maize Ubiquitin gene, Cry1A.105 nucleotide sequence, Cry2Ab nucleotide sequence, Hpt gene and Nos terminator sequence) were transferred to maize chromosome In the group, the corn plants transferred to Cry1A.105 nucleotide sequence, the corn plant transferred to Cry2Ab nucleotide sequence and the corn plant transferred to Cry1A.105-Cry2Ab nucleotide sequence were obtained; as comparison.

[0108] 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 an application of an insecticidal protein. A control method for chilotraea infuscatellus pests comprises the following step of at least contacting the chilotraea infuscatellus pests with a protein CrylA.105. The chilotraea infuscatellus pests are controlled through generating the protein CrylA.105 capable of killing chilotraea infuscatellus in a plant body. Compared with an agricultural control method, a chemical control method and a physical control method adopted in the prior art, the control method can be used for avoiding invasion of the chilotraea infuscatellus pests by protecting the whole plant in the whole growth period and is pollution-free, residue-free, stable and thorough in effect, simple, convenient and economic.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry1A.105 protein to control the plant damage caused by the pod borer by expressing it in plants. Background technique [0002] Chestnut gray borer (Chilo infuscatellus), also known as sugarcane two-spot borer, belongs to Lepidoptera moth family. It is widely distributed in Southeast Asia, South Asia and China. In China, it mainly damages gramineous crops, such as corn, sorghum, millet and sugarcane. Millet ash borer is a stem-boring pest, which can cause dead plants after the plant is damaged. Under normal circumstances, the spring valley area and the spring-summer valley mixed-seeding area have heavy occurrences, and the summer valley area has light damage. [0003] Corn is an important food crop in China. With the strengthening of the global greenhouse effect, the temperature has continued to rise in the past two years, and the types and numbers of in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01P7/04C12N15/84A01H5/00A01H1/02
CPCA01N47/44A01N63/50
Inventor 杨旭丁德荣张欣馨康越景
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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