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Application of rab11a gene and its dsRNA in migratory locust control

A technology of migratory locusts and genes, applied in the direction of DNA / RNA fragments, applications, genetic engineering, etc., can solve the problems of abnormal development, death, and various life activities of insects cannot be carried out normally, and achieve high lethality and specificity Strong, long-lasting control effect

Active Publication Date: 2022-07-19
SHANXI UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

After silencing the target gene, various life activities of insects cannot be carried out normally, resulting in abnormal development and final death of insects

Method used

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  • Application of rab11a gene and its dsRNA in migratory locust control
  • Application of rab11a gene and its dsRNA in migratory locust control

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Sequence of locust small GTPase gene 11A (LmRab11A) and acquisition of dsRNA

[0025] 1. Obtaining the locust small GTPase gene 11A (LmRab11A) sequence

[0026] 1) Search the locust Rab11A gene sequence in the locust transcriptome database

[0027] Based on the migratory locust transcriptome database, the sequence of Rab11A gene was searched by bioinformatics method. After sequence analysis and alignment, a total of 1 sequence of Rab11A gene was obtained.

[0028] 2) Design of primers required for PCR amplification:

[0029] The upstream and downstream primers were designed according to the above-mentioned gene fragments using primer premier 5.0 software. The primer sequences are shown in SEQ ID NO: 3 and SEQ ID NO: 4, respectively. The primers were synthesized by Shanghai Sangon Bioengineering Co., Ltd.

[0030] 3) PCR amplification reaction

[0031] The total RNA of the third instar nymphs was extracted, and the extracted RNA was reverse transcribed into first-stra...

Embodiment 2

[0044] Experiment of killing migratory locusts by dsRNA of small GTPase gene 11A (LmRab11A) in migratory locusts

[0045] 1. Specific dsRNA Injection

[0046] 2 μl (6 μg) of the dsRNA synthesized in the above Example 1 was injected with a 25 μl microsyringe into the second and third abdominal segments of the first-day instar nymphs of the third instar migratory locust. The control group was injected with the same volume and concentration of dsEGFP. The injected locusts were raised in a constant temperature incubation room at 30°C (light: dark time=14h:10h, temperature 30±2°C, humidity 60%), and the control and treatment groups were fed fresh wheat seedlings and wheat bran every day.

[0047] 2. Silence detection of locust small GTPase gene 11A (LmRab11A)

[0048] 9 locusts injected with dsEGFP (control group) and dsLmRab11A (experimental group) for 24 hours were collected, the whole nymphs were taken for total RNA extraction, and reverse transcribed into first-strand cDNA, a...

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Abstract

The invention relates to the field of biotechnology, in particular to the application of the locust Rab11A gene and its dsRNA in the control of migratory locusts. Specifically, the fragment of the small GTPase gene 11A is obtained from the transcriptome of the migratory locust by bioinformatics method, and after further cloning and sequencing, the Rab11 gene of the migratory locust is obtained, and the sequence is SEQ ID NO: 1. According to SEQ ID NO: 1, the dsRNA of the gene was designed and synthesized. The nucleotide sequence of one chain of the dsRNA is shown in SEQ ID NO: 2. After injecting it into the body cavity of the migratory locust, it can specifically silence the target gene, and the dsRNA can specifically silence the target gene. Locusts die when they molt, and many experiments have shown that the lethality rate reaches 100%. Due to the specificity and high lethality of the present invention, it has important practical significance for pest control, and can provide a new way for pest control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a migratory locust Rab11A gene and its dsRNA in the control of migratory locusts. Background technique [0002] Locusts are traditional agricultural pests in my country. Due to their migratory and omnivorous characteristics, once a locust plague breaks out, it will cause serious harm to agricultural economic production. The use of chemical pesticides to prevent and control locust plagues leads to pesticide residues, environmental pollution, and food safety. Long-term use will also cause locusts to develop obvious drug resistance. Therefore, the development of new efficient and environmentally friendly pesticides is an important way to solve the current situation. [0003] RNA interference (RNAi) technology is a specific post-transcriptional gene silencing phenomenon induced by double-stranded RNA (dsRNA). The phenomenon was officially named by two scientists, And...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/55C12N15/10C12N15/113C12N15/11A01N57/16A01P7/04
Inventor 李涛史学凯王艳丽范云鹤张建珍
Owner SHANXI UNIV
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