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Application 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 invasion of Spodoptera frugiperda pest, however, there is no report on the control of plant damage by Spodoptera litura 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

[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 (605 amino acids) of Cry1Fa insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Cry1Fa nucleotide sequence (1818 nucleotides) of encoding corresponding to the amino acid sequence of said Cry1Fa insecticidal protein , as shown in SEQ ID NO:4 in the sequence listing.

[0089] 2. Synthesize the above nucleotide sequence

[0090] The Cry2Ab nucleotide sequence (as shown in SEQ ID NO: 2 in the sequence listing) and the Cry1Fa nucleotide sequence (as shown in SEQ ID NO: 4 in the sequence listing) we...

no. 2 example

[0091] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0092] 1. Construction of a recombinant cloning vector containing the Cry2Ab gene

[0093] 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).

[0094] Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 c...

no. 3 example

[0103] The third embodiment, the acquisition of transgenic plants

[0104] 1. Obtaining transgenic corn plants

[0105] 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 the recombinant expression vectors DBN100033 and DBN100076 (including the promoter sequence of the CaMV35S gene, the Cry2Ab nucleotide sequence, the Cry1Fa nucleotide sequence, the Hpt gene and the Nos terminator sequence) were transferred into the maize genome, and obtained The corn plant transformed with the Cry2Ab nucleotide sequence and the corn plant transformed with the Cry2Ab-Cry1Fa nucleotide sequence; at the same time, the wild-type corn plant was used as a control.

[0106] For Agrobacterium-mediated transformation of maize, briefly, immature immature embryos are isolated from maize and contacted with a susp...

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PUM

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Abstract

The invention relates to an application of an insecticidal protein. A method for controlling prodenia litura comprises the step of enabling the prodenia litura to contact Cry2Ab protein at least. The prodenia litura is controlled and killed by Cry2Ab protein generated inside plants. Compared with agricultural control methods, chemical control methods and physical control methods in the prior art, the insecticidal protein can protect the wholes plants in the whole growth period against the attack of prodenia litura, is free from pollution and residue, is stable and thorough in effect, is simple and economical and is convenient to use.

Description

technical field [0001] The invention relates to the use of an insecticidal protein, in particular to the use of a Cry2Ab protein to control plants from being damaged by Spodoptera litura through expression in plants. Background technique [0002] Spodoptera litura belongs to Lepidoptera Noctuidae. It is an omnivorous and gluttonous pest. It damages a wide range of hosts. In addition to corn and soybeans, it can also damage melons, eggplants, beans, onions, leeks, spinach and cruciferous There are nearly 100 families and more than 300 kinds of plants such as vegetables, grains, and economic crops; Spodoptera litura is distributed worldwide, and it occurs all over the country, mainly in the Yangtze River Basin and the Yellow River Basin. Spodoptera litura mainly damages the whole plant with larvae, and gathers and eats on the back of leaves when it is young; after the third instar, it disperses and damages leaves, tender stems, and old larvae can eat fruits. [0003] Corn and...

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
CPCA01H4/00C12N15/82
Inventor 李建勇杨旭李梅于彩虹
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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