Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)

被引：20

作者：

Bai, Yu ^{[1
]}

Lv, Ya-Nan ^{[1
]}

Zeng, Mei ^{[1
]}

Jia, Pei-Yao ^{[1
]}

Lu, Hu-Na ^{[1
]}

Zhu, Yi-Bo ^{[1
]}

Li, Sheng ^{[1
]}

Cui, Ying-Ying ^{[1
]}

Luan, Yun-Xia ^{[1
]}

机构：

[1] South China Normal Univ, Inst Insect Sci & Technol, Sch Life Sci, Guangdong Prov Key Lab Insect Dev Biol & Appl Tec, Guangzhou 510631, Peoples R China

来源：

GENES | 2021年 / 12卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Thermobia domestica; reference genes; expression stability; quantitative real-time PCR; RNA interference;

D O I：

10.3390/genes12010021

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Zygentoma occupies a key evolutionary position for understanding the evolution of insect metamorphosis but has received little attention in terms of genetic analysis. To develop functional genomic studies in this insect, we evaluated five candidate internal reference genes for quantitative RT-PCR (qPCR) studies from Thermobia domestica, a representative species of Zygentoma, including Actin 5C (Actin5C), Elongation factor-1 alpha (EF1A), Ribosome protein S26 (RPS26), Ribosome protein L32 (RPL32), and Superoxide dismutase 2 (SOD2), at different developmental stages, in various body parts, and under dsRNA microinjection and starvation stresses, using four algorithms (delta Ct, geNorm, NormFinder and BestKeeper) and a comparative algorithm (RefFinder). Specific suitable reference genes were recommended across specific experimental conditions, and the combination of RPS26 and RPL32 was appropriate for all tested samples. Employing our selected reference gene combination, we investigated the gene expression pattern of Myoglianin (Myo), a crucial gene-regulating insect metamorphosis, in ametabolous T. domestica, and demonstrated the efficiency of RNA interference (RNAi) in firebrat nymphs. This study provides a basis for reliable quantitative studies of genes and greatly benefits evolutionary and functional genomics studies in Zygentoma.

引用

页码：1 / 14

页数：14

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