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Low-density lipoprotein receptor related gene 2 LmLRP2 of migratory locust and application of dsRNA of gene in prevention and control of migratory locust

A low-density lipoprotein and receptor technology, applied in DNA/RNA fragments, receptors/cell surface antigens/cell surface determinants, applications, etc., can solve environmental pollution, endanger human health, pesticide residues, etc.

Active Publication Date: 2019-08-09
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Once a locust plague occurs, it is almost impossible to stop it. At present, chemical control is mainly used to control it. However, long-term use of chemical insecticides can easily lead to a series of problems such as environmental pollution, pesticide residues, and harm to human health.

Method used

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  • Low-density lipoprotein receptor related gene 2 LmLRP2 of migratory locust and application of dsRNA of gene in prevention and control of migratory locust
  • Low-density lipoprotein receptor related gene 2 LmLRP2 of migratory locust and application of dsRNA of gene in prevention and control of migratory locust
  • Low-density lipoprotein receptor related gene 2 LmLRP2 of migratory locust and application of dsRNA of gene in prevention and control of migratory locust

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Obtaining the sequence of migratory locust low-density lipoprotein receptor-related gene 2 and its dsRNA

[0018] 1. Sequence acquisition of low-density lipoprotein receptor-related gene 2 in migratory locusts

[0019] Based on the migratory locust transcriptome database, the sequence of the migratory locust low-density lipoprotein receptor-related gene 2 was searched using bioinformatics methods, and the migratory locust low-density lipoprotein receptor-related gene 2 ( LmLRP2 ) part of the sequence. Use the NCBI website to perform Blast analysis on the sequence, and at the same time combine the genome sequence of migratory locust to obtain its coding sequence, use BioEdit software to splice and compare, and obtain LmLRP2 Full length open reading frame sequence. The upstream and downstream primers were designed using primer premier5.0 software and sent to Shanghai Sangon Bioengineering Co., Ltd. for synthesis.

[0020] Extract the total RNA of the 5th ins...

Embodiment 2

[0026] Embodiment 2: Migratory locust low-density lipoprotein receptor-related gene 2 dsRNA kills 4-year-old migratory locust

[0027] 1. Injection of low-density lipoprotein receptor-related gene 2-specific dsRNA in migratory locusts

[0028] 4 µl (10 µg) dsRNA of SEQ ID NO: 1 (ds LRP2 ) with a 25 μl micro-syringe to inject between the second and third abdominal segments of the 4th instar nymphs on the first day. A total of 30 nymphs were injected, half male and half male. Inject the same volume and concentration of ds GFP into the control group. After injection, migratory locusts were raised in a 30°C constant temperature biochemical incubator (light:dark time = 14 h: 10 h, temperature 30±2°C, humidity 40±5%), and fed fresh wheat seedlings and wheat bran every day.

[0029] 2. Detection of silencing efficiency of migratory locust low-density lipoprotein receptor-related gene 2

[0030] 24 h after dsRNA injection, the epidermis of nymphs was extracted for RNA extraction...

Embodiment 3

[0033] Example 3: Migratory locust low-density lipoprotein receptor-related gene 2 dsRNA affects the permeability of migratory locust epidermis

[0034] After 4th instar nymphs were injected with dsRNA, the molted nymphs were transferred to a 2 ml centrifuge tube containing 1.5 ml dye solution (0.5% eosin (W / V, Sigma, a red water-soluble dye with a molecular mass of 691.86 Da)), After 30 minutes at 45°C, wash with water three times. The results showed that injection of ds GFP The migratory locusts cannot be colored while injecting ds LmLRP2 The epidermis of migratory locusts showed obvious coloring ( image 3 ), indicating that the permeability of the epidermis has changed.

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Abstract

The invention provides a low-density lipoprotein receptor related gene 2 LmLRP2 of migratory locust and application of dsRNA of the gene in prevention and control of the migratory locust. The specificlow-density lipoprotein receptor related gene 2 can be silenced to kill the migratory locust. After cloning and sequencing of the low-density lipoprotein receptor related gene 2 of the migratory locust, the low-density lipoprotein receptor related gene 2 with the sequence shown as SEQ ID NO:1 is obtained; the dsRNA of the gene is designed and synthesized according to SEQ ID NO:1. After the dsRNAof the gene is injected into the body cavity of the migratory locust, the migratory locus dies before molting, during molting or after molting separately, and the permeability of the epidermis is enhanced. A new specific molecular target is provided for pest prevention and control on the basis of RNA interference, and a new approach is provided for pest prevention and control.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to low-density lipoprotein receptor-related gene 2 of migratory locust LmLRP2 and its application of dsRNA in migratory locust control. Background technique [0002] Migratory locust is an important worldwide agricultural pest with a wide distribution range and the characteristics of gathering in groups and long-distance migration. Once a locust plague occurs, it is almost impossible to stop it. At present, chemical control is mainly used to control it. However, long-term use of chemical insecticides can easily lead to a series of problems such as environmental pollution, pesticide residues, and harm to human health. Therefore, it is an important scientific issue to screen new molecular targets for green control of migratory locusts. [0003] RNA interference (RNAi) refers to the phenomenon that cells use endogenous or exogenous double-stranded RNA (dsRNA) to induce, and speci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113A01N57/16A01P7/04
CPCC07K14/705C12N15/113A01N57/16C12N2310/14Y02A50/30
Inventor 张建珍赵艺妍赵小明刘卫敏马恩波
Owner SHANXI UNIV
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