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Tea geometrid reverse transcription transposon gene rtp and application thereof

A technology of retrotransposon, tea inchworm, applied in genetic engineering, plant genetic improvement, application, etc., can solve the problem of small larvae

Inactive Publication Date: 2008-07-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that NPV has a good control effect on young tea inchworms, but has little effect on larvae above the third instar

Method used

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  • Tea geometrid reverse transcription transposon gene rtp and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] Take 10 late 1st instar larvae and 2 4th instar tea geometrid larvae, extract total RNA with TRIZOL (Invitrogen Company) reagent, and use UNIQ-10 Column mRNA Extraction and Purification Kit (Shanghai Sangon Bioengineering Co., Ltd.) Isolate mRNA. 1 μg of mRNA was used for cDNA synthesis, and double-stranded cDNA was synthesized using TakaRa MMLV RTase cDNASynthesis Kit (Takara Bioengineering [Dalian] Co., Ltd.). After the double-stranded cDNA was purified by isopropanol precipitation, it was digested with a combination of TakaRaBspHI and BssHII (Bao Bioengineering [Dalian] Co., Ltd.). For specific operations, refer to the instructions of BspHI and BssHII restriction enzymes. After adding an equal volume of isopropanol to the digested product and ice-bathed for 1 hour, centrifuge at 13,000 rpm for 25 minutes to collect the precipitate. The precipitate was mixed with two previously designed linkers, SEQ ID No: 3 and SEQ ID No: 4, and ligated with T4 DNA ligase (Shanghai ...

Embodiment 2

[0028] Dilute the NPV virus preparation with water to 10.0×10 5 PIB / ml, smear the front and back of the tea leaves with the diluted virus liquid, and feed the 1-year-old, 2-year-old, 3-year-old and 4-year-old tea geometrids after drying in the room. After 2 hours of feeding, 2 samples were taken in each treatment. -10 larvae. The above larval samples were extracted with TRIZOL (Invitrogen Company) for total RNA, and PCR was carried out with a redesigned primer combination of SEQ ID No: 14 and SEQ ID No: 15 (the expected product length was 341bp) to study the expression of rtp at different ages and the expression of larvae on Correlation of NPV viral resistance. The results showed that after the NPV virus preparation treated the larvae of the 1st instar tea geometrid, the mortality rate of the larvae was about 93%, while that of the 2nd instar larvae was about 75%; The case fatality rate of larvae after virus treatment was 18%. Gene expression analysis showed that the rtp ge...

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PUM

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Abstract

The invention discloses a tea geometrid retrotransposon gene rtp with a nucleotide sequence which is shown in SEQ ID No: 1. The invention also discloses the encoding protein of the gene rtp with the nucleotide sequence which is shown in SEQ ID NO: 2. The invention also discloses the use of the gene rtp: the gene is utilized to regulate the anti-virus properties of lepidoptera pests.

Description

technical field [0001] The invention belongs to the field of insect genetic engineering. Specifically, the present invention relates to the isolation and analysis of a tea geometrid retrotransposon (rtp) gene; and also involves using the gene to regulate the tea geometrid's ability to resist viruses. Background technique [0002] Tea inchworm is one of the main pests in tea gardens, so it is also an important control object. With the pollution-free and organic management of tea gardens, the use of chemically synthesized pesticides has been restricted or banned, while biological pesticides have become vigorously promoted preparations. Nuclear polyhedrosis virus (NPV) is a biopesticide commonly used to control tea geometrids. Studies have shown that NPV has a good control effect on young tea geometrids, but has little effect on larvae above the third instar. The exploration of the reason why the tea geometrid produces resistance to NPV will help the rational and efficient u...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435A01K67/033
Inventor 陆建良林晨梁月荣董占波郑新强
Owner ZHEJIANG UNIV
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