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Use of insecticidal protein

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

Active Publication Date: 2018-10-19
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 gene can resist the damage of Lepidoptera pests such as corn borer and millet borer. However, there is no report about controlling the damage of plants by hornworm by producing transgenic plants expressing Cry2Ab protein.

Method used

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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 (1178 amino acids) of Cry1Ac insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Cry1Ac nucleotide sequence (3537 nucleotides) of encoding corresponding to the amino acid sequence of said Cry1Ac insecticidal protein , as shown in SEQ ID NO:4 in the sequence listing.

[0089] The amino acid sequence (1177 amino acids) of Cry1A.105 insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1A.105 nucleotide sequence ( 3534 nucleotides), as shown in SEQ ID NO:6 in the sequence list...

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 Cry2Ab gene

[0094] 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).

[0095] Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 co...

no. 3 example

[0106] The third embodiment, the acquisition of transgenic plants

[0107] 1. Obtaining transgenic soybean plants

[0108] According to the routinely used Agrobacterium infection method, the cotyledon node tissue of the aseptically cultivated soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 3 in the second example, so that the recombinant gene constructed in 2 in the second example T-DNA of expression vectors DBN100033, DBN100075 and DBN100076 (including promoter sequence prAtUbi10 of Arabidopsis ubiquitin gene, Cry2Ab nucleotide sequence, Cry1Ac nucleotide sequence, Cry1A.105 nucleotide sequence, PAT gene and tNos termination subsequence) into the soybean genome, obtained the soybean plant transferred to the Cry2Ab nucleotide sequence, the soybean plant transferred to the Cry2Ab-Cry1Ac nucleotide sequence and the soybean transferred to the Cry2Ab-Cry1A.105 nucleotide sequence plants, and wild-type soybean plants were used as controls.

[010...

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PUM

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Abstract

The invention relates to use of a pesticidal protein, and the use of the pesticidal protein comprises contacting Clanos bilineata pests at least with Cry2Ab protein. The Clanos bilineata pests can becontrolled by production of the Cry2Ab protein capable of killing Clanos bilineata in a plant; compared with the agricultural control method, the chemical control method, the physical control method and the biological control method used in the prior art, the whole plant can be protected in the whole growth period for control of damage of the Clanos bilineata pests, and the pesticidal protein is non-pollution, free of residue, stable in effect, 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 Cry2Ab protein expressed in plants to control the bean hornworm from injuring plants. Background technique [0002] Bean horn moth Clanis bilineata (walker), its larvae are commonly known as bean worm, bean dan, bean cicada, belongs to the genus Bean horn moth of Lepidoptera Scutellaridae Scutellaria subfamily, mainly distributed in the Huanghuai River Basin and the Yangtze River Basin and South China, the main soybean producing area in my country, is one of the main soybean pests. The bean hornworm is an explosive pest, and its damage characteristics are: the larvae overeat the leaves, the light ones are bitten into holes and nicks, and the severe ones become polished stems, enter the residual petioles, and cannot set pods, seriously affecting the yield. [0003] Cultivated soybean (Glycine max (L.) Merri) is an important economic crop grown globally as ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H6/54A01N37/46A01P7/04A01H1/02A01H4/00A01H1/06
Inventor 任振涛吴竹筠韩超杨淑靖
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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