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A kind of plant anti-insect protein and its coding gene and application

An insect-resistant gene and plant technology, applied in the application field of plant insect resistance, can solve problems such as hidden dangers of human and animal safety, pesticide residues, ecological environment impact, etc., and achieve the effect of good insecticidal effect and good resistance.

Active Publication Date: 2021-08-06
LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, chemical control methods are mainly used for the control of Spodoptera frugiperda at present, and the insect has developed strong resistance to many pesticides
In addition, problems such as pesticide residues caused by the use of chemical control methods have hidden dangers to human and animal safety and risks to the ecological environment.

Method used

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  • A kind of plant anti-insect protein and its coding gene and application
  • A kind of plant anti-insect protein and its coding gene and application
  • A kind of plant anti-insect protein and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The acquisition and synthesis of embodiment 1Cry1Fa gene

[0029] 1. Obtain the Cry1Fa nucleotide sequence

[0030] The amino acid sequence (605 amino acids) of mCry1Fa insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; The mCry1Fa nucleotide sequence (1818 nucleotides), as shown in SEQ ID NO:1 in the sequence listing.

[0031] The amino acid sequence (605 amino acids) of Cry1Fa insecticidal protein, as shown in SEQ ID NO:4 in the sequence listing; The Cry1Fa nucleotide sequence (1818 nucleotides), as shown in SEQ ID NO:2 in the sequence listing.

[0032] 2. Synthesize the above-mentioned Cry1Fa nucleotide sequence

[0033] The mCry1Fa nucleotide sequence (as shown in SEQ ID NO: 1 in the sequence listing) and the Cry1Fa nucleotide sequence (as shown in SEQ ID NO: 3 in the sequence listing) were synthesized by Nanjing KingScript Biotechnology Co., Ltd. The 5' end of the synthetic mCry1Fa nucleotide sequence is also connected with an NcoI restriction...

Embodiment 2

[0038] Example 2 Construction of a recombinant expression vector containing the Cry1Fa gene

[0039] Recombinant cloning vector LP05-T and expression vector LP-BB (vector backbone: pCAMBIA3301 (CAMBIA organization can provide)) were respectively digested with restriction endonucleases NcoI and EcoRI, and the excised mCry1Fa nucleotide sequence fragment was inserted into the expression Between the NcoI and EcoRI sites of the vector LP-BB, it is well known to those skilled in the art to construct the vector by using the conventional enzyme digestion method. The recombinant expression vector LP-PT05 is constructed, and its construction process is as follows: figure 2 Shown (Kan: kanamycin gene; RB: right border; Ubi: maize Ubiquitin (ubiquitin) gene promoter (SEQ ID NO: 8); mCry1Fa: mCry1Fa nucleotide sequence (SEQ ID NO: 1); Nos: the terminator of nopaline synthase (SEQ ID NO: 6); Ubi: the maize Ubiquitin (ubiquitin) gene promoter (SEQ ID NO: 8); PAT: the gene encoding phosphin...

Embodiment 3

[0042] Embodiment 3 recombinant expression vector transforms Agrobacterium

[0043] The correctly constructed recombinant expression vectors LP-PT05 and LP-PT05-CK were transformed into Agrobacterium LBA4404 (Invitrgen, Chicago, USA; Cat.No: 18313-015) with liquid nitrogen method, and the transformation conditions were: 100 μL Agrobacterium LBA4404, 3 μL plasmid DNA (recombinant expression vector); placed in liquid nitrogen for 10 minutes, 37 ° C warm water bath for 10 minutes; inoculate the transformed Agrobacterium LBA4404 in LB test tubes at a temperature of 28 ° C and a rotation speed of 200 rpm Cultivate for 2 hours, spread on LB plates containing 50 mg / L of Rifampicin and 50 mg / L of Kanamycin until a positive single clone grows, pick a single clone to culture and extract its plasmid, The recombinant expression vectors LP-PT05 and LP-PT05-CK were digested with restriction endonucleases NotI and SalI, and the results showed that the structures of the recombinant expression...

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Abstract

The present application discloses a protein for plant anti-insect, the amino acid sequence of the protein is SEQ ID NO:3. At the same time, the coding gene SEQ ID NO: 1 of the insect-resistant protein is also provided. The present invention further provides the application of the above-mentioned insect-resistant protein and gene in resisting cutworms. The insect-resistant protein and gene provided by the invention can provide better cutworm resistance and fall armyworm resistance.

Description

technical field [0001] This application relates to the technical field of genetic engineering biological control, in particular to the artificially modified insect-resistant gene mCry1Fa, its coded and expressed protein, and its application in plant insect resistance. Background technique [0002] Biological control is the use of certain beneficial organisms or biological metabolites to control the population of pests to achieve the purpose of reducing or eliminating pests, such as Trichogramma or Beauveria bassiana to control meadow borers. It is characterized by being safe for humans and animals, less polluting to the environment, and can achieve long-term control of some pests; however, the effect is often unstable, and the same investment is required regardless of the severity of meadow moth occurrence. In order to solve the limitations in the practical application of agricultural control, chemical control, physical control and biological control, scientists have discove...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8286
Inventor 贾志伟李晓娇吕玉平刘枫王强李树秀孙宇李涛赵丽媛张原
Owner LONGPING BIOTECHNOLOGY (HAINAN) CO LTD
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