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Sequence of locust I type chitinase gene, and application of dsRNA of locust I chitinase gene

A chitinase gene and locust technology, applied in the field of sequence and its dsRNA application, can solve the problems of endangering human health, drug resistance of locusts, pesticide residues, etc.

Inactive Publication Date: 2014-07-23
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Pests are the enemy of my country's agricultural production. Long-term use of chemical pesticides has led to a series of problems such as pesticide residues, serious environmental pollution, drug resistance of locusts, and harm to human health.

Method used

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  • Sequence of locust I type chitinase gene, and application of dsRNA of locust I chitinase gene
  • Sequence of locust I type chitinase gene, and application of dsRNA of locust I chitinase gene
  • Sequence of locust I type chitinase gene, and application of dsRNA of locust I chitinase gene

Examples

Experimental program
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Effect test

Embodiment 1

[0011] Example 1: The sequence of migratory locust type I chitinase gene and its dsRNA

[0012] 1. Obtaining the sequence of migratory locust type I chitinase gene

[0013] 1) Searching the sequence of migratory locust type Ⅰ chitinase gene in migratory locust transcriptome database

[0014] Based on the migratory locust transcriptome database, a bioinformatics method was used to search for the sequence of migratory locust type I chitinase gene. After sequence analysis and comparison, a total of 1 migratory locust type I chitinase gene sequence was obtained Fragment, the cDNA sequence lacks the 3'end.

[0015] 2) Obtain the full-length cDNA sequence of migratory locust type I chitinase gene

[0016] Based on the obtained fragments of migratory locust type I chitinase gene, primer premier 5.0 software was used to design 3'RACE primers. All primers were synthesized by Shanghai Yingweijieji Biological Co., Ltd. The total RNA of 5th instar nymphs of migratory locust was extracted, and th...

Embodiment 2

[0022] Example 2: Migratory locust type I chitinase gene dsRNA killed 5th instar migratory locust

[0023] 1. Injection of locust type Ⅰ chitinase gene dsRNA

[0024] 1μl (3μg) of SEQ ID NO: 2 dsRNA was injected with a 25μl micro-syringe into the second and third abdominal segments of the second-day-old nymphs of the 5th instar migratory locust. A total of 40 nymphs, half male and half male, were injected. The same volume concentration of dsGFP was injected into the control group. After injection, the migratory locusts were kept in a 30°C constant temperature biochemical incubator.

[0025] 2. Observation of the phenotype of 5th instar migratory locust after injection of dsRNA

[0026] After the 5th instar nymphs were injected with dsRNA, the control group injected with dsGFP began to molt 6 days later and they all successfully moulted to adults. The average moulting time was 15-20 minutes. After moulting to adults, the morphological activity of the nymphs was normal, and they began ...

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Abstract

The invention provides a sequence of locust I type chitinase gene (cht5), and an application of dsRNA of the locust I chitinase gene. The dsRNA can silence the I type chitinase gene to make locusts dead. The I type chitinase gene with the sequence represented by SEQ ID NO:1 is obtained by cloning the locust I type chitinase gene, and sequencing; and a gene fragment with the sequence represented by SEQ ID NO:2 is selected to synthesize double strand RAN (dsRNA). After the dsRNA is injected to the body cavity of the locust, the locust has delayed growth, and cannot successfully molt to enter a next age period, so the locust is dead. A new target is provided for the RNA interference based insect control in the invention.

Description

Technical field [0001] The invention relates to the field of biotechnology. Specifically, it relates to the sequence of locust type I chitinase gene and the application of dsRNA. Background technique [0002] Pests are the enemy of my country's agricultural production. The long-term application of chemical pesticides has caused a series of problems such as pesticide residues, serious environmental pollution, resistance of locusts to pesticides, and endangering human health. With the development of society, there is an urgent need for new pest control methods. [0003] In recent years, pest control technology based on RNA interference (RNAi) is developing rapidly. This method has the following advantages: 1) It chooses to interfere with specific genes of pests, which is safe for higher animals and humans; 2) It is specific to insects and has no killing effect on non-target organisms; 3) It is not harmful to the environment. Toxic and harmless. [0004] The pest control program is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N15/113A01N61/00A01P7/04
Inventor 李大琪张建珍马恩波孙毅
Owner SHANXI UNIV
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