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The application of insecticidal protein

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

Active Publication Date: 2017-03-15
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 Cry1A gene can resist the invasion of Lepidoptera pests such as cotton bollworm and fall armyworm. However, there is no research on the production of transgenic plants expressing Cry1A protein to control the damage of armyworm to plants. report

Method used

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  • The application of insecticidal protein
  • The application of insecticidal protein
  • The application of insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0082] The first embodiment, the acquisition and synthesis of genes

[0083] 1. Obtain the nucleotide sequence

[0084] The amino acid sequence (818 amino acids) of Cry1Ab-01 insecticidal protein, as shown in SEQ ID NO:1 in the sequence listing; Cry1Ab-01 nucleotide sequence ( 2457 nucleotides), as shown in SEQ ID NO:5 in the sequence listing.

[0085] The amino acid sequence (615 amino acids) of Cry1Ab-02 insecticidal protein, as shown in SEQ ID NO: 2 in the sequence listing; Cry1Ab-02 nucleotide sequence ( 1848 nucleotides), as shown in SEQ ID NO:6 in the sequence listing.

[0086] 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:7 in the sequence listing.

[0087] The amino acid sequence (1178 amino acids) of the Cry1Ac insecticidal protein, as shown in the sequence table SEQ ID NO: 4; the Cry1Ac nucleotide sequence (3537...

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 Cry1A gene

[0094] The synthesized Cry1Ab-01 nucleotide sequence was connected to 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 , its construction process is as follows 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; Cry1Ab-01 is the Cry1Ab-01 nucleotide sequence (SEQ ID NO:5); MCS is the multiple cloning site).

[0095]Then, the recombinant cloning vector DBN01-T was transformed into Escheric...

no. 3 example

[0112] The third embodiment, the acquisition of transgenic corn plants

[0113] 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 transformed with the recombinant expression vector described in 3 in the second embodiment, so that the second embodiment T-DNA in the recombinant expression vectors DBN100738, DBN100053, DBN100106, DBN100029, DBN100076 and DBN100734 constructed in Example 2 (comprising the promoter sequence of the corn Ubiquitin gene, Cry1Ab-01 nucleotide sequence, Cry1Ab-02 nucleotide sequence, Cry1A.105 nucleotide sequence, Cry1Ac nucleotide sequence, Vip3A nucleotide sequence, Cry2Ab nucleotide sequence, PMI gene and Nos terminator sequence) were transferred into the maize genome, and the Cry1Ab-01 nuclear The corn plant of nucleotide sequence, the corn plant of Cry1Ab-02 nucleotide sequence, the corn plant of Cry1Ab-01-Vip3A nucleotid...

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Abstract

The invention relates to the application of an insecticidal protein. The method for controlling mythimna lorgi pests comprises the steps of making mythimna lorgi contact with at least Cry1A protein. The application method uses the fact that a plant body can produce Cry1A protein which kills mythimna lorgi to control the mythimna lorgi pests. Compared with the prior arts of agricultural control method, chemical control method and physical control method, the application method can achieve a protection range covering the full breeding season and the whole plant to prevent the infiltration of mythimna lorgi. The application has the advantages of having no pollution and no residue, being stable and complete in effect, and being simple, convenient and economical.

Description

technical field [0001] The invention relates to an application of an insecticidal protein, in particular to an application of a Cry1A protein expressed in plants to control damage to plants by Armyworm rougei. Background technique [0002] Mythimna seperata belongs to Lepidoptera Noctuidae. It is an omnivorous, migratory and intermittently explosive pest. It mainly feeds on corn, wheat, sorghum, millet, rice, soybean and other crops. It is distributed in Henan, Shandong, Jiangsu, Anhui, Hunan, Hubei and Guangdong and Guangxi regions of China. In Henan and Shandong areas, there are 3-4 generations of Myxoma lauii a year, and the overwintering adults are seen from mid-April to early May, and end in mid-October. It mainly harms spring corn. [0003] Corn is an important food crop in China. With the strengthening of the global greenhouse effect, the temperature has continued to rise in the past two years, and the types and numbers of insect pests have increased. Due to the sma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/02A01G13/00A01N47/44A01P7/04A01H5/10
CPCA01G13/00A01H1/02A01N47/44C07K14/325C12N15/8286
Inventor 丁德荣张爱红
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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