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Bactrocera dorsalis eth gene and its ligand, receptor and its application in fruit fly specific control agent

A kind of tangerine fruit fly, ETH-2-R technology, applied in the field of genetic engineering, to achieve the effect of good application prospects

Inactive Publication Date: 2019-12-10
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, GPCRs in insects are rarely reported as pesticide targets, and the application in Bactrocera dorsalis is even less.

Method used

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  • Bactrocera dorsalis eth gene and its ligand, receptor and its application in fruit fly specific control agent
  • Bactrocera dorsalis eth gene and its ligand, receptor and its application in fruit fly specific control agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The acquisition of the open reading frame sequence of embodiment 1 Bacteralis dorsalis ETH and its receptor

[0038] 1. Primer design and amplification of the open reading frame sequence of Bactrocera dorsalis ETH and its receptor:

[0039] Based on the Bactrocera dorsalis genome and transcriptome database, the ETH and ETH-R-A, ETH-R-B of Drosophila melanogaster were used for TBLASTN search, and the ETH and its receptors (ETH-R-A and ETH-R-B) of Bactrocera dorsalis were predicted. The nucleotide sequence of R-B) and, the nest PCR specific primer of design amplification ETH is ETH-1-F / ETH-1-R and ETH-2-F / ETH-2-R, amplifies ETH-R-A The primers for amplification and amplification of ETH-R-B are ETH-R-A-F / ETH-R-A-R, and the primers for amplifying ETH-R-B are ETH-R-B-F / ETH-R-B-R. The primer sequences are as follows:

[0040]

[0041] The total RNA of B. dorsalis was extracted with an RNA extraction kit, and reverse transcribed into cDNA with a reverse transcription kit. ...

Embodiment 2

[0053] The measured open reading frame sequence of the Bactrocera dorsalis ETH receptor ETH-R-A is shown in SEQ ID NO:14 in the sequence listing, and the open reading frame sequence of the receptor ETH-R-B is shown in SEQ ID NO:15 in the sequence listing . Example 2: Determination of the binding ability of the ligand polypeptide to its receptor based on the detection of intracellular calcium ion flow

[0054] 1. Construction of CHO (Chinese hamster ovary cell) cell expression plasmids of Bactrocera dorsalis ETH-R-A and ETH-R-B

[0055] Construction of pcDNA3.1-ETH-R-A expression vector: Add Kpn I and XbaI restriction sites to the ORF of ETH-R-A by PCR method, and use Kpn I and XbaI to carry out enzyme digestion, pcDNA3.1(+) plasmid , also after double digestion with Kpn I and Xba I, ligation was carried out, and after ligation, transformation was carried out by referring to the above-mentioned transformation method. After the positive identification of the bacterial liquid wa...

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Abstract

The invention discloses a citrus fruit fly ETH gene. The sequence of the citrus fruit fly ETH gene is represented by SEQ ID NO:9. The invention further discloses an amplification method of the citrus fruit fly ETH gene. According to the amplification method, nest type PCR is adopted, and primers are ETH-1-F / ETH-1-R and ETH-2-F / ETH-2-R. The invention further discloses a citrus fruit fly ETH ligand. The amino acid sequence of the citrus fruit fly ETH ligand is represented by SEQ ID NO:12 or 13, and nucleotide sequences for encoding the amino acid sequence are respectively represented by SEQ ID NO:10-11. The invention further discloses an open reading frame sequence of a citrus fruit fly ETH receptor represented by SEQ ID NO:14 or 15. The invention further discloses an amplification method of the citrus fruit fly ETH receptor. According to the amplification method, primer pairs ETH-R-A-F / ETH-R-A-R and ETH-R-B-F / ETH-R-B-R are adopted. The invention further discloses application of the gene, the ligand and the receptor in neuroregulation of flies.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a Bactrocera dorsalis ETH gene, its ligand, receptor and its application in a specific control agent for Bactrocera dorsalis. Background technique [0002] Citrus is an important economic crop. With the adjustment of my country's agricultural industry structure, the scale of citrus industry has continued to expand, and it has gradually become a pillar industry in many regions. Bactrocera dorsalis is a serious threat to many economically important fruits and vegetables, especially citrus, due to its strong environmental adaptability and ability to invade and spread. At present, Bactrocera dorsalis has become a huge threat to the healthy development of the citrus industry. At present, Bactrocera dorsalis has developed severe resistance to pesticides such as organophosphorus insecticides, pyrethroids, and avermectins on the market; pesticide residues seriously pollute...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/16C12N15/10C07K7/08C12N15/11C12N15/12
CPCC07K7/08C07K14/575C07K14/723C12N15/1096C12Q2531/113C12Q2549/119
Inventor 王进军石岩蒋红波刘天远王磊豆威
Owner SOUTHWEST UNIV
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