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Whole-set reagent and method for inhibiting gene expression of pine wood nematode

A pine wood nematode and gene expression technology, applied in the biological field, can solve problems such as high cost and inability to continuously interfere

Active Publication Date: 2017-05-31
CHINA AGRI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

RNA interference technology (RNAi) has been widely used in the study of the gene function of plant parasitic nematodes. The soaking method is more common when applying RNAi technology to study the gene function of pine xylophilus, but it is expensive and cannot be continuously interfered.

Method used

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  • Whole-set reagent and method for inhibiting gene expression of pine wood nematode
  • Whole-set reagent and method for inhibiting gene expression of pine wood nematode
  • Whole-set reagent and method for inhibiting gene expression of pine wood nematode

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Embodiment 1

[0066] Example 1. A complete set of reagents for inhibiting gene expression of pine xylophilus and its application in reducing high temperature sensitivity and locomotor activity of pine xylophilus

[0067] The set of reagents provided in this example for inhibiting gene expression of pine xylophilus consists of pDE3dBH.qa vector, pBT6 plasmid and Neurospora crassa strain dcl-1 - / dcl-2 - Composition, this embodiment utilizes pDE3dBH.qa vector, pBT6 plasmid and Neurospora crassa strain dcl-1 - / dcl-2 - The expression of target genes of pine xylophilus is suppressed, and the selected target genes are hsp gene, cyc gene and tmy gene. Specific steps are as follows:

[0068] 1. Construction of dsRNA expression vector

[0069] Synthesize AC fragment (hsp-AC) and BC fragment (hsp-BC) for suppressing hsp gene expression, the sequences from 5' end to 3' end of hsp-AC and hsp-BC are respectively sequence 1 and sequence in the sequence listing 2. Wherein, the 1st-6th of the sequen...

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Abstract

The invention discloses a whole-set reagent and a method for inhibiting gene expression of pine wood nematode. The whole-set reagent comprises a whole-set reagent 1 and a whole-set reagent 2, wherein the whole-set reagent 1 comprises a whole-set reagent A and nucleic acid molecules reducing expression of hsp gene, the whole-set reagent A comprises a pDE3dBH.qa carrier, a neurospora crassa dcl-1- / dcl-2 and pBT6 plasmid, and the whole-set reagent 2 comprises a whole-set reagent A and nucleic acid molecules reducing expression of cyc gene or nucleic acid molecules reducing expression of tmy gene. Experiments prove that the expression of target genes (hsp, tmy and cyc genes) of pine wood nematode can be successfully inhibited by utilizing the whole-set reagent and the method, and further high-temperature resistance sensitivity can be reduced and the movement activity can be inhibited.

Description

technical field [0001] The invention relates to a set of reagents and a method for inhibiting gene expression of pine xylophilus in the field of biotechnology. Background technique [0002] Pine xylophilus is a quarantine pathogen at home and abroad, and with its wide spread, it poses a serious threat to pine forests in my country. RNA interference technology (RNAi) has been widely used in the study of gene function of plant parasitic nematodes. Soaking method is more common when applying RNAi technology to study gene function of pine xylophilus, but it is expensive and cannot be continuously interfered. There is an urgent need for a simple method for interfering with gene expression in B. xylophilus. Contents of the invention [0003] The technical problem to be solved by the invention is how to reduce the expression of the target gene of the pine xylophilus and how to reduce the high temperature sensitivity of the pine xylophilus and inhibit the movement activity of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/80C12N1/15A01N63/04A01P5/00
CPCA01K67/033A01N63/30C12N15/80
Inventor 刘倩简恒王梦陈永攀
Owner CHINA AGRI UNIV
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