Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
Autor: | Pei-Yao Jia, Hu-Na Lu, Yu Bai, Ying-Ying Cui, Yun-Xia Luan, Ya-Nan Lv, Sheng Li, Mei Zeng, Yi-Bo Zhu |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: |
0301 basic medicine
Genetics biology lcsh:QH426-470 Lepismatidae reference genes Thermobia domestica biology.organism_classification RNA interference 03 medical and health sciences lcsh:Genetics 030104 developmental biology 0302 clinical medicine expression stability Reference genes Gene expression Thermobia quantitative real-time PCR Firebrat Functional genomics Gene 030217 neurology & neurosurgery Genetics (clinical) |
Zdroj: | Genes, Vol 12, Iss 21, p 21 (2021) Genes Volume 12 Issue 1 |
ISSN: | 2073-4425 |
Popis: | 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. |
Databáze: | OpenAIRE |
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