Application of insecticidal protein

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

Active Publication Date: 2019-02-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] It has been proved that the plants transgenic for Cry2Ab can resist the damage of Lepidoptera pests such as corn borer and millet borer. However, there is no report about controlling the damage of meadow borer 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

[0087] The first embodiment, the acquisition and synthesis of genes

[0088] 1. Obtain the nucleotide sequence

[0089] 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.

[0090] 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.

[0091] 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

[0095] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0096] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0097] 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).

[0098]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 co...

no. 3 example

[0116] The third embodiment, the acquisition of transgenic plants

[0117] 1. Obtaining transgenic soybean plants

[0118] According to the commonly used Agrobacterium infection method, the cotyledon node tissue of the aseptically cultivated soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 4 in the second embodiment, so that the soybean constructed in 2 in the second embodiment T-DNA of recombinant expression vectors DBN100033, DBN100175, DBN100176 and DBN100003 (including Cry2Ab nucleotide sequence, Cry1Ac nucleotide sequence, Cry1A.105 nucleotide sequence, Vip3A nucleotide sequence and PAT gene) were transferred to soybean chromosome In the group, soybean plants transferred to Cry2Ab nucleotide sequence, soybean plants transferred to Cry1Ac-Cry2Ab nucleotide sequence, soybean plants transferred to Cry1A.105-Cry2Ab nucleotide sequence and Vip3A nucleotide sequence were obtained sequenced soybean plants, and wild-type soybean plants were used ...

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Abstract

The invention relates to application of insecticidal protein. The application comprises the step of enabling meadow moth pests to at least contact Cry2Ab protein. Through generation of the Cry2Ab protein capable of killing meadow moths in bodies of plants, the meadow moth pests are controlled; compared with an agricultural control method, a chemical control method, a physical control method and abiological control method in the prior art, the invention has the advantages as follows: the whole plants are protected in the whole growth period so as to control invasion of the meadow moth pests, and no pollution, no residue, a stable and thorough effect, simpleness, convenience and economy are achieved.

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 a plant to control the plant damage caused by the meadow borer. Background technique [0002] The meadow moth Loxostege sticticalis, belonging to the Lepidoptera Boreridae, is mainly distributed in the Northeast, Northwest, and North China, namely Jilin, Inner Mongolia, Heilongjiang, Ningxia, Gansu, Qinghai, Hebei, Shanxi, Shaanxi, Jiangsu and other provinces . Meadow borer is a polyphagous large pest, which can feed on 35 families and more than 200 kinds of plants, mainly sugar beets, soybeans, sunflowers, potatoes, hemp, vegetables, medicinal materials and other crops; suffer from it. The meadow moth usually has a major outbreak every 10 years or so. The characteristics of its damage are: the larvae mostly gather on the branches and form nets to hide, eat the mesophyll, and leave the epidermis and veins. In severe cas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/54A01N37/46A01P7/04A01H1/02A01H4/00A01H1/06
CPCA01H1/02A01H1/06A01H4/008A01N37/46C12N15/8286
Inventor 张爱红杨淑靖任振涛吴竹筠
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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