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Use of insecticidal protein

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

Active Publication Date: 2015-05-20
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 peach borer to plants by producing transgenic plants expressing Cry2Ab protein.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0084] The first embodiment, the acquisition and synthesis of genes

[0085] 1. Obtain the nucleotide sequence

[0086] 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.

[0087] 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.

[0088] 2. Synthesize the above nucleotide sequence

[0089] 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

[0090] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0091] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0092] 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).

[0093] Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 c...

no. 3 example

[0102] The third embodiment, the acquisition of transgenic plants

[0103] 1. Obtaining transgenic corn plants

[0104] 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.

[0105] For Agrobacterium-mediated maize transformation, briefly, immature immature embryos were isolated from maize and contacted with a suspension of Agrobacterium capable of trans...

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Abstract

The invention relates to the use of insecticidal protein. A method for controlling dichocrocis punctiferalis pests comprises the step of at least contacting the dichocrocis punctiferalis pests with Cry2Ab protein. According to the use of the insecticidal protein, the dichocrocis punctiferalis pests can be controlled by the Cry2Ab protein which is capable of killing the dichocrocis punctiferalis and is generated in plants; compared with the agricultural control method, chemical control method and biological control method which are used in the prior art, the use has the advantages that the insecticidal protein can be used for protecting the whole plants in the whole growth period so as to control the harm of the dichocrocis punctiferalis pests, and is free from pollution and residue, stable and thorough in effect, simple, convenient and economical.

Description

technical field [0001] The invention relates to the use of an insecticidal protein, in particular to the use of a Cry2Ab protein expressed in plants to control the damage of plants by the peach borer borer. Background technique [0002] Peach borer (Conogethes punctiferalis) belongs to the family Lepidoptera and is an omnivorous pest. It not only harms corn, sorghum and other crops, but also harms peaches, persimmons, chestnuts and other fruit trees. It is widely distributed in my country, starting from Heilongjiang and Inner Mongolia in the north. , south to Taiwan, Hainan, Guangdong, Guangxi, and the southern edge of Yunnan, east to the former Soviet Union, North Korea, west from Shanxi, Shaanxi to Ningxia, Gansu, and then to Sichuan, Yunnan, and Tibet. When maize is harmed, it mainly eats ears and stems, and the plant damage rate reaches 30%-80%. When sorghum is harmed, newly hatched larvae eat into young sorghum grains and seal their mouths with feces or food residues. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N47/44A01P7/04C12N15/84A01H5/00A01H1/02A01H4/00A01G13/00A01G1/00
Inventor 李建勇李梅庞洁
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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