Internal reference gene capable of stable expression in different tissues of Sogatella furcifera, and screening method and application thereof

Abstract

The invention relates to an internal reference gene capable of stable expression in different tissues of Sogatella furcifera, and a screening method and application thereof, belonging to the technical field of biology. The nucleotide sequence of the internal reference gene RPL9 is disclosed as SEQ ID NO:7. The method comprises the following steps: designing degenerate primers according to other insect related gene sequences disclosed in NCBI, carrying out PCR (polymerase chain reaction) to obtain 6 common internal reference gene partial nucleotide sequences of Sogatella furcifera, and carrying out cloning, sequencing and NCBI database Blast comparison to determine the partial sequences of the Sogatella furcifera internal reference gene; and designing specific quantitative primers on the basis of the partial sequences, establishing an RT-qPCR (reverse transcription-quantitative polymerase chain reaction) process based on an SYBR Green I dye technique, and screening out the internal reference gene RPL9 capable of stable expression in different tissue parts of Sogatella furcifera by using a fluorescent quantitative PCR technique. The internal reference gene lays solid foundation for researching gene expression level of different tissue parts and gene functions of Sogatella furcifera in 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 a screening method and application of internal reference genes stably expressed in different tissue parts of white-backed planthopper. 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. The white-backed planthopper is widely distributed in my country, including South China, Central China, North China, Northeast China, and Southwest China. The northwest rice area can reach Miquan in Xinjiang Autonomous Region and other places. Rice is a suitable host for white-backed planthopper. White-backed planthopper can not only suck rice phloem juice, but al...

AI Technical Summary

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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