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DsRNA and application thereof to German cockroach

A German cockroach, DNA molecule technology, applied in the field of genetic engineering, can solve problems such as spoilage, food taste, and chewing clothes

Active Publication Date: 2018-11-27
MEIZHOU HUASHENG GUANGMEI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although German cockroaches are small in size, they are extremely lethal. They bite bad clothes, contaminate food, and spread diseases. They are carriers of various bacteria; German cockroaches can secrete some strange smell substances. When the population density is high , these secretions will fill their living places with a special smell, and at the same time, these secretions can make food taste bad and deteriorate, and bring certain harm to human life and agricultural production

Method used

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  • DsRNA and application thereof to German cockroach
  • DsRNA and application thereof to German cockroach

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Experimental program
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Embodiment 1

[0021] Synthesis of embodiment 1 dsRNA

[0022] Total RNA was extracted from the fat body of female Blattella germanica using TRIzol (Life technologies), and then the first cDNA strand was synthesized by reverse transcription (PrimeScript II reverse transcriptase (Takara Bio, Shiga, Japan)) using oligodT primers.

[0023] According to the existing genome information of the German cockroach, the complete sequence of the Tet2 gene of the German cockroach was found by comparing with other species, as shown in SEQ ID No.1.

[0024] Use the dsRNA design website E-RNAi (https: / / www.dkfz.de / signaling / e-rnai3 / ), copy and paste the entire targeted open reading frame sequence of the German cockroach Tet2 gene into the website, and screen through the design parameters Get the best dsRNA targeting sequence. Then design primers: Tet2F: CAGCGTGGATTGTGGTAG (SEQ ID No.2) and Tet2R: AATGGTCGTCCTGGTTTG (SEQ ID No.3). Using cDNA as a template to amplify the DNA fragment containing the target s...

Embodiment 2

[0026] The German cockroach living injection experiment of embodiment 2 dsRNA

[0027] Female German cockroaches that have just emerged into adults were selected and mixed with male adults. On the second day after eclosion, they were anesthetized at low temperature and placed on a microscope dissection table, and then microinjected Tet2-targeted The dsRNA of the gene was injected into the abdomen of the German cockroach according to different doses. Then repeat every two days, and inject three times in total. After injection, continue to observe the effect of dsRNA treatment on the oviposition of female Blattella germanica, and study the effect of Tet2 gene on the reproduction and oviposition of Blattella germanica when the protein expression level is reduced.

[0028] (1) Validation of dsRNA interference effect on Tet2 gene

[0029] 48 hours after the two injections of dsRNA, the German cockroach was anesthetized and dissected, the fat body was dissected out and RNA was extr...

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Abstract

The invention discloses dsRNA and application thereof to German cockroach. DsRNA which targets a Tet2 gene of German cockroach is double-stranded DNA consisting of a nucleotide with a nucleotide sequence represented by SEQ ID No.7 and a nucleotide reversely complemented with a nucleotide represented by SEQ ID No.7. DsRNA is synthesized by virtue of an RNAi interference technique and is used for targeting a demethylated relevant gene Tet2 of DNA, and therefore, the development of ovaries of cockroaches is influenced, and the prevention and control of the cockroaches are realized based on the blockage or damage of the development of the ovaries and the formation of oothecae.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a dsRNA and its application in preventing and treating the German cockroach. Background technique [0002] Cockroaches are native to Africa and live in a humid, warm, and dark place; they are small and like to hide in hidden crevices; their activities are concentrated at night, so it is difficult for humans to eliminate their populations. [0003] German cockroach is a relatively common cockroach. It is an omnivorous insect. It can eat sour, sweet, bitter, spicy and salty food, insect corpses, etc., and prefers to eat sweets. Food and water are crucial to the survival of German cockroaches. Research by Wang Xuejun and others has shown that German cockroaches have a strong ability to withstand hunger. Under starvation, German cockroaches will eat each other. Water is more important than food to the survival of German cockroaches. If there is food and water, adults c...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/10A01N57/16A01P7/04
CPCA01N57/16C12N15/113C12N2310/14C12N2330/30
Inventor 任充华李胜
Owner MEIZHOU HUASHENG GUANGMEI BIOTECHNOLOGY CO LTD
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