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A kind of dsrna of locust fatty acid synthase gene lmfas and preparation method and application

A technology of fatty acid synthase and gene, which is applied in the field of genetic engineering, can solve problems such as ecological environment impact, environmental pollution, crop hazards, etc., and achieve the effect of broad application prospects

Active Publication Date: 2022-03-18
杨青
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Migratory locust is an important agricultural pest with strong jumping and migratory ability, which makes it have a strong ability to survive and adapt to the environment, thus causing great harm to crops
At present, the control of agricultural pests mainly relies on chemical pesticide control. Although this method is fast, efficient and stable, it has disadvantages that cannot be ignored: 1) pollutes the environment, threatens human health and affects other non-target organisms 2) The emergence of insecticide resistance will reduce or even eliminate the control efficiency, which will have a huge impact on the ecological environment

Method used

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  • A kind of dsrna of locust fatty acid synthase gene lmfas and preparation method and application
  • A kind of dsrna of locust fatty acid synthase gene lmfas and preparation method and application
  • A kind of dsrna of locust fatty acid synthase gene lmfas and preparation method and application

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Experimental program
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Effect test

Embodiment 1

[0028] A dsRNA of migratory locust fatty acid synthase gene LmFAS, the nucleotide sequence of the dsRNA is SEQ ID NO:2.

[0029] A method for preparing the dsRNA of migratory locust fatty acid synthase gene LmFAS, comprising the following steps:

[0030] a, Based on the migratory locust transcriptome database, the bioinformatics method was used to search the migratory locust fatty acid synthase gene. After sequence analysis, splicing and comparison, the LmFAS sequence of the migratory locust fatty acid synthase gene was obtained, and the upstream primer sequence and downstream Primer sequences, and synthesized;

[0031] b, Choose five-instar migratory locust nymphs that grow healthy and have equal size, half male and half male, and quickly dissect their epidermis under a stereo microscope, and freeze them in liquid nitrogen. Three first biological repetitions were set up, and four biological repetitions were set up, and RNA was extracted according to the Trizol kit of TaKaRa ...

Embodiment 2

[0036] Lethal test of migratory locust fatty acid synthase gene dsRNA:

[0037] 1. dsRNA injection of migratory locust fatty acid synthase gene

[0038]Thirty 4th-instar 1st-day nymphs and 5th-instar 1st-day nymphs of healthy growth, uniform size, and half male and half male were selected for the experiment. Gently inject 10 μg of synthetic dsLmFAS into the flank of the nymph between the second and third abdominal segments along the bloodstream using a 25 μl microsyringe. At the same time, 30 nymphs were selected as the control group. Inject the same volume and concentration of dsGFP into the control group. The injected migratory locusts were reared in a constant temperature biochemical incubator at 30°C (light:dark time=14h:10h, temperature 30±2°C, humidity 60%), and fed fresh wheat seedlings and wheat bran every day.

[0039] 2. Gene silencing detection of migratory locust fatty acid synthase

[0040] Nine nymphs injected with dsGFP and dsLmFAS were collected, and total ...

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Abstract

The invention belongs to the technical field of genetic engineering, and in particular relates to a dsRNA of migratory locust fatty acid synthase gene LmFAS, a preparation method and application thereof. The preparation method of the dsRNA of migratory locust fatty acid synthase gene LmFAS of the present invention, comprises the following steps: a, design upstream primer sequence and downstream primer sequence, and synthesize; B, extract RNA and reverse transcribe into first-strand cDNA as The template, combined with the upstream and downstream primers of the fatty acid synthase gene LmFAS of migratory locusts, was amplified by PCR to obtain the full-length fragment of the fatty acid elongase gene, the obtained product was purified, connected to the T4 vector, and then transformed into Trans1‑T1 competent cells Medium culture, spot picking, detection, and sequencing; c, based on the fatty acid synthase gene sequence SEQ ID NO: 1 of migratory locust, design dsRNA upstream and downstream primers, the sequences of which are SEQ ID NO: 3 and SEQ ID NO: 4, the upper Downstream primers all carry T7 promoter sequence; d, dsRNA is transcribed and synthesized.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a dsRNA of migratory locust fatty acid synthase gene LmFAS, a preparation method and application thereof. Background technique [0002] Migratory locust is an important agricultural pest, which has a strong ability to bounce and migrate, which in turn makes it have a strong ability to survive and adapt to the environment, thus causing great harm to crops. At present, the control of agricultural pests mainly relies on chemical pesticide control. Although this method is fast, efficient and stable, it has disadvantages that cannot be ignored: 1) pollutes the environment, threatens human health and affects other non-target organisms 2) The emergence of insecticide resistance will reduce or even eliminate the control efficiency, which will have a huge impact on the ecological environment. Therefore, there is an urgent need to develop novel green pest control m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/10A01N57/16A01P7/04
CPCC12N15/1137A01N57/16C12N2310/14C12Y203/01085C12N2330/30
Inventor 赵小明张建珍
Owner 杨青
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