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Conopomorpha sinensis bradley vitellogenin gene CsVg, coding protein and application thereof

A litchi pedicle borer and vitellogenin technology is applied in the fields of application, genetic engineering, and plant genetic improvement. It can solve the problems of detection methods that have no effect on fecundity, and achieve short detection cycles, improved sensitivity and accuracy, and stable good sex effect

Active Publication Date: 2018-12-18
PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in actual production, there is no detection method that can be used for the impact of this agent on the fecundity of litchi pedicle borer, and this problem needs to be solved urgently.

Method used

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  • Conopomorpha sinensis bradley vitellogenin gene CsVg, coding protein and application thereof
  • Conopomorpha sinensis bradley vitellogenin gene CsVg, coding protein and application thereof
  • Conopomorpha sinensis bradley vitellogenin gene CsVg, coding protein and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] The cDNA cloning method of litchi pedicle borer vitellogenin gene CsVg comprises the following steps:

[0026] (1) The total RNA was extracted from the female adult moth of the litchi pedicle borer, and the extracted total RNA was used as a template to synthesize the first strand of cDNA by reverse transcription; then the cDNA template of the litchi pedicle borer was synthesized by PCR.

[0027] (2) Design degenerate primers CsVg-F: 5'-CMYMTGADGTNMGRGTKGCNGC-3' and CsVg-R: 5'-GCASWRTANGGCCANACRC-3' according to the existing Lepidoptera insect Vg conserved region, see SEQ ID NO.3 and SEQ ID NO.4, (where R=A / G, Y=C / T, M=A / C, K=G / T, S=C / G, W=A / T, H=A / C / T, B=C / G / T, V=A / C / G, D=A / G / T, N=A / C / G / T).

[0028] (3) performing PCR amplification with the cDNA template described in step (1) and the degenerate primers described in step (2), and purifying the obtained PCR product.

[0029] (4) Perform agarose gel electrophoresis on the PCR product purified in step (3), recover the tar...

Embodiment 2

[0036] Utilize the full-length CsVg vitellogenin gene CsVg of litchi pedicle borer provided by the present invention, and establish a kind of molecular detection method that beta-cypermethrin affects the litchi pedicle borer's fecundity by means of real-time fluorescent quantitative PCR technology, the molecular detection method comprises the following steps:

[0037] (1) Taking the vitellogenin gene CsVg of the litchi pedicle borer as the target sequence, according to the principle of real-time fluorescent quantitative PCR primer design, design Real-CsVg-F:5'-ATGCAACCATAGAGTCGC-3' and Real-CsVg-R:5'-ATCATCTTCCGTAGATACCAG -Two pairs of 3' specific detection primers, see SEQ ID NO.7 and SEQ ID NO.8.

[0038] (2) Total RNA samples were extracted from the surviving litchi pedicle borer female adults in the treatment group (30ppm concentration of lambda-cypermethrin spraying treatment) and the litchi pedicle borer female adults in the control group (normal feeding), and the total R...

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Abstract

The invention discloses a conopomorpha sinensis bradley vitellogenin gene CsVg, a coding protein and application thereof. The nucleotide sequence of the CsVg gene is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein thereof is shown as SEQ ID NO.2. The invention also discloses a detection method for detecting the influence of the drug on conopomorpha sinensis bradley fecundity, the method detects the expression quantity of the vitellogenin gene CsVg of conopomorpha sinensis Bradley that are applied with and not applied with the drug to judge whether the conopomorpha sinensis bradley fecundity is affected after drug application, has the advantages of high detection sensitivity and accuracy, short detection cycle, good stability, etc., can be used for high-throughput detection of the influence of the drug on conopomorpha sinensis bradley fecundity, and provides molecular reference basis for rational use of the drug in actual production of litchis and longans.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the vitellogenin gene CsVg, a key factor of litchi pedicle moth fecundity, and its encoded protein and application. Background technique [0002] Vitellogenin (Vg) gene is a protein specifically present in non-mammalian female animals, which plays a decisive role in the maturation and development of insect fertilized eggs. So far, the Vg genes of various insects have been successfully cloned and identified, but there is no report about Vg in the family Mothidae. The present invention discovers the vitellogenin gene CsVg in the Litchi pedicle borer, which is a major pest in the litchi and longan industry for the first time, and analyzes the amino acid sequence encoded by it, and supplements the information on the molecular characteristics of the vitellogenin gene in the Litchiidae insects . [0003] Litchi and longan are the famous and precious fruits of our country, and the cultiva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12Q1/6888C12N15/10
CPCC07K14/43563C12N15/10C12Q1/6888
Inventor 姚琼董易之阙引利徐淑陈炳旭
Owner PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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