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Migratory locust wing specific cuticle protein gene and application of dsRNA thereof

A kind of epidermal protein and specific technology, applied in DNA/RNA fragments, applications, genetic engineering, etc., can solve problems such as pesticide residues, serious environmental pollution, and harm to human health

Inactive Publication Date: 2016-07-13
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term application of chemical pesticides leads to a series of problems such as pesticide residues, serious environmental pollution, insect resistance, and harm to human health.

Method used

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  • Migratory locust wing specific cuticle protein gene and application of dsRNA thereof
  • Migratory locust wing specific cuticle protein gene and application of dsRNA thereof
  • Migratory locust wing specific cuticle protein gene and application of dsRNA thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0012] Example 1: The sequence, tissue expression and dsRNA acquisition of migratory locust wing epidermal protein gene

[0013] 1. Obtaining the sequence of the wing epidermal protein gene of migratory locust and its tissue expression analysis

[0014] 1) Search for the sequence of the migratory locust wing epidermal protein gene in the migratory locust transcriptome database

[0015] Based on the migratory locust transcriptome database, the sequence of the migratory locust wing epidermal protein gene was searched using bioinformatics methods. After sequence analysis and comparison, the sequence fragments of the migratory locust wing epidermal protein gene were obtained.

[0016] 2) Acquisition of the full-length cDNA sequence of the wing epidermal protein gene of migratory locust

[0017] Based on the obtained locust wing epidermal protein gene sequence, 5' RACE primers were designed using primer premier 5.0 software. All primers were synthesized by Sangon Bioengineering (...

Embodiment 2

[0025] Example 2: Migratory locust wing-specific epidermal protein gene dsRNA causes abnormal wing development of adults

[0026] 1. Injection of locust wing-specific epidermal protein gene dsRNA

[0027] 5 μl (10 μg) of dsRNA of SEQ ID NO: 2 was injected with a 25 μl micro-syringe between the second and third abdominal segments of the 2nd-day-old nymphs of the 5th instar migratory locust, and a total of 60 were injected, half male and half male. Inject dsGFP at the same volume concentration into the control group. The injected migratory locusts were raised in a constant temperature biochemical incubator at 30°C.

[0028] 2. Observation of the phenotype of migratory locust wing tissue after dsRNA injection

[0029] After the 5th instar nymphs were injected with dsRNA, 6 days after the injection of dsGFP the control group began to molt and all of them successfully molted to adults. After molting to adults, the nymphs were normal in shape and vitality, and began to eat normall...

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Abstract

The invention provides a migratory locust wing specific cuticle protein gene and application of dsRNA thereof. DsRNA of the gene can silence the specific cuticle protein gene, so that migratory locust wings grow abnormally and migratory locust finally die.Concretely, through cloning and sequencing migratory locust cuticle protein gene, the wing cuticle protein gene with the sequence being SEQ ID NO:1 is obtained, and RT-PCR and western blot results show specific existence of the wing cuticle protein gene in migratory locust wing tissue; then a gene segment with the sequence being SEQ ID NO:2 is selected and used for double-stranded RNA (dsRNA) synthesis. After the gene dsRNA is injected into body cavities of migratory locust nymphs, wings of adult nymphs grow abnormally after molting, are manifested to grow with malformation and shrinkage and finally the migratory locusts die.A new target is provided for insect prevention and treatment based on RNA interference.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of locust wing-specific epidermal protein gene and dsRNA. Background technique [0002] Pests are the enemy of agricultural production in our country. For some harmful insects, because they have wings, they can migrate and fly long distances between lands, thus greatly improving their efficiency in foraging, courtship, avoiding enemies, and expanding distribution. , this habit has become the main reason why some pests cause disasters in different places. The long-term application of chemical pesticides has led to a series of problems such as pesticide residues, serious environmental pollution, insect resistance, and harm to human health. With the development of society, new pest control methods are urgently needed. [0003] In recent years, pest control technology based on RNA interference (RNAi) is developing rapidly. The method has the following advantages: 1) It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113A01N57/16A01P7/04
CPCC07K14/43563A01N57/16C12N15/113C12N2310/10
Inventor 赵小明勾昕秦忠玉马恩波张建珍
Owner SHANXI UNIV
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