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Application of insecticidal proteins

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

Active Publication Date: 2019-05-28
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 for Cry2Ab can resist damage from Lepidoptera (Lepidoptera) pests such as corn borer and pomegranate moth. However, there is still no research on controlling the damage to plants by producing transgenic plants expressing Cry2Ab protein. to report

Method used

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  • Application of insecticidal proteins
  • Application of insecticidal proteins
  • Application of insecticidal proteins

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; 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 the Cry1A.105 insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; the Cry1A.105 nucleotide sequence ( 3534 nucleotides), as shown in SEQ ID NO:6 in the sequence ...

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 of 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; flori 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 compete...

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 embodiment, so that the soybean constructed in 2 in the second embodiment The T-DNA of recombinant expression vectors DBN100033, DBN100175 and DBN100176 (including Cry2Ab nucleotide sequence, Cry1Ac nucleotide sequence, Cry1A.105 nucleotide sequence and PAT gene) were transferred into the soybean genome, and the transferred Cry2Ab The soybean plant of the nucleotide sequence, the soybean plant of the Cry1Ac-Cry2Ab nucleotide sequence and the soybean plant of the Cry1A.105-Cry2Ab nucleotide sequence are used as the control.

[0109]For Agrobacterium-mediated soybean transformation, briefly, mature soybean seeds were germinated in soybean germ...

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Abstract

The invention relates to an application of insecticidal proteins. The application of the insecticidal proteins comprises the following steps that argyrogramma agnata pests are at least in contact withCry2Ab proteins. According to the application of the insecticidal proteins, the Cry2Ab proteins capable of killing argyrogramma agnata are generated in plant bodies to control the argyrogramma agnatapests. Compared with an agricultural control method, a chemical control method, a physical control method and a biological control method used in the prior art, full-growth period and full-plant protection on plants are adopted in the application of the insecticidal proteins to control the invasion of the argyrogramma agnata pests, and the application of the insecticidal proteins has no pollutionand residue, has stable and thorough effects, and is 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 Cry2Ab protein to control plant damage to plants by expressing in plants. Background technique [0002] Argyrogramma agnata, belonging to Lepidoptera Noctuidae, is mainly distributed in the Yangtze River Basin and the Yellow River Basin in my country. It is one of the main soybean pests in the main soybean producing areas of China. Silver print moth is a polyphagous pest, mainly harming beans, rapeseed, cabbage, cauliflower, cabbage, radish and other cruciferous vegetables and other crops. Eat up the leaves and affect the yield. Cultivated soybean (Glycine max (L.) Merri) is an important economic crop grown globally as the main source of vegetable oil and vegetable protein, and is an important food crop in China. Soybean is one of the plants that silver-leaved moths like to feed on. Every year, silver-leafed moths will cause different degrees of food lo...

Claims

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Application Information

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IPC IPC(8): A01G13/00A01G22/15A01G22/40A01H1/02A01H4/00A01H5/00A01H6/54C12N15/32C12N15/82
Inventor 陶青杨淑靖任振涛王强
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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