dsRNA, coding gene, preparation method and application thereof designed based on sex pheromone receptor gene or5m of Periplaneta americana

A technology of Periplaneta americana and receptor gene, applied in the field of dsRNA, can solve problems such as climbing, drug residues, threats to human and animal health and the ecological environment, and achieve the effects of controlling the number and speed of reproduction, blocking activity, and being environmentally friendly

Active Publication Date: 2021-09-28
SOUTH CHINA NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the control of Periplaneta americana still mainly relies on traditional chemical pesticides, but problems such as drug resistance, pesticide residues, and re-emergence are increasing, seriously threatening the health of humans and animals and the ecological environment

Method used

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  • dsRNA, coding gene, preparation method and application thereof designed based on sex pheromone receptor gene or5m of Periplaneta americana
  • dsRNA, coding gene, preparation method and application thereof designed based on sex pheromone receptor gene or5m of Periplaneta americana
  • dsRNA, coding gene, preparation method and application thereof designed based on sex pheromone receptor gene or5m of Periplaneta americana

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Design and synthesis of dsRNA

[0045] The design and synthesis of dsRNA includes the following steps:

[0046] (1) Primer design

[0047] From the reported OR5M gene sequences of other species, a homologous alignment was performed in the Periplaneta americana genome to obtain the complete sequence of the Periplaneta americana sex pheromone receptor gene OR5M (as shown in SEQ ID No.1). Using the dsRNA design website E-RNAi (https: / / www.dkfz.de / signaling / e-rnai3 / ), copy and paste the entire Periplaneta americana sex pheromone receptor gene OR5M open reading frame sequence into the website, and design The dsRNA targeting sequence I (as shown in SEQID No.2) was obtained by parameter screening. Primers were designed according to the nucleotide sequence shown in SEQ ID No.2, forward primer OR5M-Fp: ATTGCACGGAATTTGTT (SEQ ID No.5) and reverse primer OR5M-Rp: AGTCTGAGTAGTGGACTG (SEQ ID No.6).

[0048] The OR5M sequence (SEQ ID No.1) of the sex pheromone receptor ...

Embodiment 2

[0067] Example 2 The expression level of OR5M in different tissues and different developmental times of Periplaneta americana

[0068] Fluorescent quantitative PCR primers were designed using primer premier5 primer design software. The primers used for fluorescent quantitative PCR detection are shown in Table 1. SYBR Green qPCR mix (yisheng, China) was used for quantitative detection of gene expression, and antennae (male and female), mouthparts, forefoot, brain, wings, The expression level of OR5M gene in testis and ovary (such as figure 1 shown), and the expression level of the OR5M gene in male antennae within 1-9 days after eclosion was further determined (such as figure 2 shown).

[0069] From figure 1 It can be seen that the expression level of OR5M in male antennae is much higher than that in other tissues, and the expression level of male antennae is significantly higher than that of female antennae, which is consistent with the characteristics that sex pheromones ...

Embodiment 3

[0072] Embodiment 3dsRNA influences on OR5M gene interference

[0073] (1) Live injection of the dsRNA that specifically inhibits the sex pheromone receptor gene OR5M of the American cockroach

[0074] Select healthy male and female Periplaneta americana that have just completed eclosion, raise them individually, and divide them into an experimental group and a control group (injection of dsGFP that cannot target any gene of the American cockroach). On the third day after eclosion, the males were anesthetized with carbon dioxide without damage, and then placed on the dissecting table, and a total of 2 μg of dsRNA targeting the OR5M gene was injected along the third to third abdomen of the abdomen by microinjection. The 4 abdominal segments (from bottom to top) were slowly injected into the abdominal cavity. At the same time, the same dose of GFP gene interference fragment was injected as a negative control. The male Periplaneta americana that has been injected with dsRNA is ...

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Abstract

The invention discloses a dsRNA designed based on the Periplaneta americana sex pheromone receptor gene OR5M, a coding gene and its preparation method and application. The nucleotide sequence of the gene is shown in SEQ ID NO.1, by synthesizing specific dsRNA To target the sex pheromone receptor gene OR5M to interfere with the recognition of the sex pheromone released by the female of the American cockroach by male adults, so as to block the normal mating of the American cockroach and finally achieve the purpose of preventing and controlling the American cockroach. The control of Periplaneta americana provides new technologies and strategies.

Description

technical field [0001] The invention relates to the field of sanitary pest control, in particular to a dsRNA designed based on the Periplaneta americana sex pheromone receptor gene OR5M, a coding gene, a preparation method and application thereof. Background technique [0002] Cockroaches are a class of important worldwide sanitary pests, causing serious harm to human health and public health. Cockroaches carry a variety of pathogenic microorganisms, which can cause infectious diseases such as cholera, anthrax, and tuberculosis, as well as allergic reactions such as skin rashes and asthma. The American cockroach (Periplaneta americana L.) is one of the most common species of cockroaches in the order Blattata, and it is also the dominant population in South China. It has super environmental adaptability and vitality, as well as amazing fecundity and detoxification ability. Periplaneta americana has developed into the dominant species of urban cockroach communities in my coun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/113A01N57/16A01P7/04
CPCA01N57/16C07K14/47C12N15/113C12N2310/14
Inventor 李娜李胜黄润董任科
Owner SOUTH CHINA NORMAL UNIVERSITY
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