Tea geometrid vitellogenin gene vtg and uses thereof

A technology of vitellogenin and tea looper, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., and can solve the problems that gene expression regulation technology has not yet been reported.

Inactive Publication Date: 2008-07-23
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although there have been many studies on the function of vtg in fish and crustaceans, there is no research report on the vtg coding gene and gene expression regulation technology of important pests of tea plants such as tea geometrid

Method used

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  • Tea geometrid vitellogenin gene vtg and uses thereof

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Effect test

Embodiment 1

[0023] Take 2-3 female adults (about 0.2 g) of the newly eclovened tea geometrid adults, put them in a mortar and grind them quickly with liquid nitrogen, add 1ml TRIZOL (Invitrogen Company) to mix well, and extract total RNA according to the steps in the TRIZOL reagent instructions . According to spectrophotometric analysis, the OD260 / OD280 ratio of the RNA sample was 1.8-2.0, and the concentration was about 850 μg / mL. Using 2 μL of the above RNA solution as a template, the first-strand cDNA was synthesized according to the instructions provided by the MMLV First-Strand cDNA Synthesis Kit (Shanghai Sangon Bioengineering Co., Ltd.).

[0024] Design and commission Shanghai Sangon Biological Products Co., Ltd. to synthesize primers with sequences of SEQ ID No: 3 (upstream primer) and SEQ ID No: 4 (downstream primer), using the first-strand cDNA synthesized above as a template, according to Table 1 The PCR is performed after adding various reactants shown in the first column, an...

Embodiment 2

[0028] Based on the nucleotide sequence of SEQ ID No: 1, TaKaRa siRNA Cocktail Kit (si-RNAase III TM ) (Bao Biological Engineering [Dalian] Co., Ltd.) to prepare vtg-siRNAs that can interact with the endogenous vtg of the tea geometrid. The prepared vtg-siRNAs were dissolved in water into an aqueous solution, and the above-mentioned aqueous solution was injected into the abdomen of the fifth-instar larvae, pupae, and female adults of the day of emergence of the tea geometrid with a microsyringe, and the injection dose was 10 ng of vtg-siRNAs per head of the tea geometrid, and The injection of pure water was used as the control. After 2 days of normal feeding, total RNA was extracted with TRIZOL (Invitrogen Company) reagent. Using SEQ ID No: 5 and SEQ ID No: 6 as primers, using the total RNA extracted from the injection-treated tea geometrid larvae, pupae and adults as templates, and performing RT-PCR with the MMLV one-step RT-PCR amplification kit, For specific operations, r...

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Abstract

The invention discloses a tea geometrid vitellogenin gene vtg, which has the nucleotide sequence shown by SEQ ID No.: 1. The invention also discloses a vtg gene encoded protein, which has the amino acid sequence shown by SEQ ID NO: 2. The invention also discloses an application of the vtg gene: the vtg gene can be used to control the reproductive capacity of female Lepidoptera pests.

Description

technical field [0001] The invention belongs to the field of insect genetic engineering. Specifically, the invention relates to a clone of a tea geometrid vitellogenin (vtg) gene; and also relates to using the gene to regulate the reproductive ability of the tea geometrid insect. Background technique [0002] Tea geometrid is one of the important pests of tea trees that commonly occurs in tea regions across the country, and its frequent occurrence often causes serious reduction in tea yield and quality. Therefore, tea geometrid is an important object of pest control in tea gardens. Although conventional pyrethroids and organophosphorus pesticides can kill tea geometrid larvae well, they have less effect on prepupa, pupae and adults; Conventional pesticides have become limited or prohibited inputs in tea gardens. Therefore, there is an urgent need to develop some new biopesticides that are nontoxic to humans and animals and can effectively control pests such as tea geometr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12Q1/68
Inventor 陆建良林晨梁月荣杜颖颖郑新强杨晓利
Owner ZHEJIANG UNIV
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