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Reference genes stably expressing sogatella furcifera at different ages, screening method and application thereof

A technology of white-backed planthopper and internal reference gene, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., and can solve problems such as expression changes

Inactive Publication Date: 2015-09-30
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Problems solved by technology

Commonly used internal reference genes are genes necessary to maintain normal cell life activities, such as Actin, 18S, GAPDH, Tublin, etc., but the mRNA expression level of no internal reference gene is invariable under all conditions. Under certain conditions, its expression may change

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  • Reference genes stably expressing sogatella furcifera at different ages, screening method and application thereof
  • Reference genes stably expressing sogatella furcifera at different ages, screening method and application thereof
  • Reference genes stably expressing sogatella furcifera at different ages, screening method and application thereof

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Embodiment Construction

[0041] The specific implementation method of the present invention is further described below, wherein the relevant operations related to molecular biology are carried out with reference to Sambrook J, Russell DW "Molecular Cloning Experiment Guide" (Science Press, third edition, translated by Huang Peitang, 2002) The primer synthesis was completed by Shanghai Sangon Bioengineering Co., Ltd., and the quantitative PCR was carried out on the ABI 7500 Real-Time PCR System in accordance with the instrument operation guide and the instructions for the use of fluorescent quantitative reagents.

[0042] 1. Breeding of white-backed planthopper source. The insect sources used in this experiment are all from the white-backed planthopper population raised in the artificial climate chamber of our laboratory. The feeding conditions are temperature: 27±1°C; humidity: 75±5%; photoperiod: 16h light: 8h dark. Tiller rice was fed with Taichung Native1 (TN1).

[0043] Collection of white-backed...

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Abstract

The invention relates to reference genes stably expressing sogatella furcifera at different ages, and a screening method and application thereof and belongs to the technical field of biology. The nucleotide sequence of the reference genes A1TU is shown as SEQ ID NO:2. A degenerate primer is designed according to the other insect related gene sequence having already been disclosed by NCBI, a PCR is used for obtaining a part of the nucleotide sequence of six frequently-used reference genes of the sogatella furcifera, and the nucleotide sequence is determined to be a part of the sequence of the reference genes of the sogatella furcifera by means of cloning, sequencing and comparing with NCBI database Blast; soon afterwards, a specific quantitative primer is designed on the basis of the sequence, an RT-qPCR method based on the SYBR Green I dye technology is set up, and the fluorogenic quantitative PCR technology is utilized for screening the most stable reference genes A1TU of the sogatella furcifera at the different ages, so that the solid basis is provided for studying the expression levels of the genes at the different ages and setting up the gene function of the sogatella furcifera in the future.

Description

technical field [0001] The invention belongs to the technical field of internal reference genes stably expressed in insects in molecular biology, and in particular relates to internal reference genes stably expressed in different ages of white-backed planthopper and its screening method and application. Background technique [0002] White-backed planthopper (Sogatella furcifera (Horváth)), belongs to the phylum Arthropoda, class Insecta, order Hemiptera, and family Planthopper. It goes through three developmental stages in its life, including egg stage, nymph stage and adult stage. Adult Has migratory habits. Rice is a suitable host for white-backed planthopper. White-backed planthopper can not only suck rice phloem juice, but also spread a variety of viral diseases, causing heavy losses to rice production. The white-backed planthopper is widely distributed in my country, including South China, Central China, North China, Northeast China, and Southwest China. The northwest ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/68
Inventor 刘玉娣安兴奎侯茂林
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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