Fusion protein for improving insect resistance of plants and application thereof

A fusion protein, plant lectin technology, applied in the field of new recombinant anti-insect fusion protein and its preparation method and use, the field of genetic engineering technology platform for anti-insect fusion protein, can solve the problem of not developing a satisfactory anti-insect protein, etc. question

Inactive Publication Date: 2015-04-29
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, to date, no satisfactory and effective insect-resistant proteins have been developed

Method used

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  • Fusion protein for improving insect resistance of plants and application thereof
  • Fusion protein for improving insect resistance of plants and application thereof
  • Fusion protein for improving insect resistance of plants and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0149] Example 1 Construction of p1301GNA, p1301AaIT and p1301AaIT / GNA (pAG) vectors

[0150] 1. Construction of p1301GNA

[0151] The intermediate vector pBluKSM-GNA was synthesized by Jerry Biological Company, including Kpn I and Sac I restriction sites. The GNA fragment was excised with Kpn I and Sac I restriction sites and connected to the vector pCAMBIA1301 to obtain p1301GNA. Gs is the signal peptide of GNA, and Gm is the mature peptide of GNA.

[0152] 2. Build p1301AaIT

[0153] The intermediate vector pBluKSM-AaIT was purchased from Jerry Biological Company, which contains two enzyme cutting sites, Kpn I and Sac I. The AaIT fragment was excised with Kpn I and Sac I restriction sites and then ligated into the vector pCAMBIA1301. The AaIT fragment was excised with Kpn I and Sac I restriction sites and connected to the vector pCAMBIA1301 to obtain p1301AaIT.

[0154] 3. Construction of p1301AaIT / GNA(pAG)

[0155] The full-length AaIT cDNA was amplified by PCR metho...

Embodiment 2

[0160] Example 2 Utilize p1301GNA, p1301AaIT and p1301AaIT / GNA (pAG) vectors to construct experimental plants

[0161] Using the plant transformation method in the general method of the present invention, the p1301GNA and p1301AaIT vectors prepared in Example 1 were transferred into Agrobacterium tumefaciens strain EHA105, and respectively transformed into tobacco, Arabidopsis and rice to obtain transgenic tobacco, Arabidopsis and rice. The obtained GNA gene transgenic plants and AaIT transgenic plants of the three crops were used as control plants in subsequent examples.

[0162] Using the plant transformation method in the general method of the present invention, the p1301AaIT / GNA (pAG) vector is transferred into the Agrobacterium tumefaciens strain EHA105, and respectively transformed into tobacco, Arabidopsis and rice to obtain the AG transgenic tobacco to be tested , transgenic Arabidopsis, transgenic rice.

[0163] The above-mentioned transgenic plants were detected, a...

Embodiment 3

[0167] The biological test of embodiment 3 cotton bollworm to transgenic tobacco

[0168] Get 30 3rd instar cotton bollworms with the same growth status, put them into the cages of the transgenic tobacco made in the above-mentioned embodiment 2 and three groups of contrast tobaccos (wild type, GNA, AaIT) respectively, and observe the growth of tobacco after 1 week . Experimental results such as Figure 8 It was shown that after 7 days after the 3rd instar cotton bollworm was released into the experimental plants, the damage of the transgenic plant AG was significantly smaller than that of other experimental types, and the resistance of the AG-transformed tobacco to the cotton bollworm was significantly improved.

[0169] Get 10 3rd instar cotton bollworms with consistent growth status and put them into the petri dish of the transgenic tobacco leaves and contrast tobacco (wild type, GNA, AaIT) leaves obtained through the above-mentioned embodiment 2, and observe the tobacco le...

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Abstract

The invention provides fusion protein for improving insect resistance of plants and an application thereof. Specifically, the invention provides fusion protein for improving insect resistance of plants. The fusion protein comprises the following structure: a neuropeptide AaIT element and a phytolectin element. The fusion protein can effectively improve the insect resistance of transgenic plants, particularly the resistance to piercing-sucking insects.

Description

technical field [0001] The invention relates to the field of biomedicine. More specifically, the present invention relates to a novel recombinant anti-insect fusion protein and its preparation method and application. The invention also relates to a genetic engineering technology platform for producing a novel recombined insect-resistant fusion protein and its application. Background technique [0002] Insect damage is an important factor affecting crop yield and quality. According to the estimates of the Food and Agriculture Organization of the United Nations, the annual loss of food due to diseases, insect pests and weeds accounts for about one-third of the total output of the world, among which the loss of insect pests is the largest, reaching 14%. There are more than 1,500 kinds of major insect pests to crops in my country, and dozens of species such as cotton bollworm, corn borer, diamondback moth, planthopper, aphid, and locust are particularly harmful. They frequent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/82C12N5/10C12N1/21C12N1/19C12N1/15
CPCY02A40/146
Inventor 李胜刘淑敏
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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