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

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 (q...

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Autores principales: Bai, Yu, Lv, Ya-Nan, Zeng, Mei, Jia, Pei-Yao, Lu, Hu-Na, Zhu, Yi-Bo, Li, Sheng, Cui, Ying-Ying, Luan, Yun-Xia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7823838/
https://www.ncbi.nlm.nih.gov/pubmed/33375665
http://dx.doi.org/10.3390/genes12010021
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author Bai, Yu
Lv, Ya-Nan
Zeng, Mei
Jia, Pei-Yao
Lu, Hu-Na
Zhu, Yi-Bo
Li, Sheng
Cui, Ying-Ying
Luan, Yun-Xia
author_facet Bai, Yu
Lv, Ya-Nan
Zeng, Mei
Jia, Pei-Yao
Lu, Hu-Na
Zhu, Yi-Bo
Li, Sheng
Cui, Ying-Ying
Luan, Yun-Xia
author_sort Bai, Yu
collection PubMed
description 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.
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spelling pubmed-78238382021-01-24 Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae) Bai, Yu Lv, Ya-Nan Zeng, Mei Jia, Pei-Yao Lu, Hu-Na Zhu, Yi-Bo Li, Sheng Cui, Ying-Ying Luan, Yun-Xia Genes (Basel) Article 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. MDPI 2020-12-25 /pmc/articles/PMC7823838/ /pubmed/33375665 http://dx.doi.org/10.3390/genes12010021 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bai, Yu
Lv, Ya-Nan
Zeng, Mei
Jia, Pei-Yao
Lu, Hu-Na
Zhu, Yi-Bo
Li, Sheng
Cui, Ying-Ying
Luan, Yun-Xia
Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title_full Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title_fullStr Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title_full_unstemmed Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title_short Selection of Reference Genes for Normalization of Gene Expression in Thermobia domestica (Insecta: Zygentoma: Lepismatidae)
title_sort selection of reference genes for normalization of gene expression in thermobia domestica (insecta: zygentoma: lepismatidae)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7823838/
https://www.ncbi.nlm.nih.gov/pubmed/33375665
http://dx.doi.org/10.3390/genes12010021
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