Systematic Selection of Suitable Reference Genes for Quantitative Real-time PCR Normalization Studies of Gene Expression in Lutjanus Erythropterus

Overview

Journal Sci Rep

Specialty Science

Date 2024 Jun 10

PMID 38858385

Authors

Lujun Chen

Qiulu Liang

Zhuoxin Lai

Haitao Cui

Zhenmin Xu

Zizhao Chen

Zhongdian Dong

Zhongduo Wang

Yusong Guo

Affiliations

Soon will be listed here.

Abstract

Quantitative real-time PCR (qRT-PCR) has been widely employed for the study of gene expression in fish, and accurate normalization is crucial. In this study, we aimed to identify the most stably expressed genes in various tissues, different developmental stages, and within astaxanthin treatment groups in Lutjanus erythropterus. Twelve candidate genes (EEF1A, CYB5R3, DLD, IDH3A, MRPL17, MRPL43, NDUFS7, PABPC1, PAGR1, PFDN2, PSMC3, and RAB10) were examined via qRT-PCR. We employed geNorm and NormFinder to assess their stability. The results revealed that RAB10 and PFDN2 exhibited relatively stable expression patterns across different tissue and astaxanthin treatment groups, while NDUFS7 and MRPL17 proved to be the most reliable reference gene combinations across various developmental stages. The stability of these selected genes was further validated by assessing the expression of two target genes, CRADD and CAPNS1, across developmental stages, reinforcing the reliability of NDUFS7 as it closely aligned with transcriptome-wide expression patterns at these stages. The present results will help researchers to obtain more accurate results in future qRT-PCR analysis in L. erythropterus.

Citing Articles

Evaluation of reference genes for gene expression analysis in Japanese flounder (Paralichthys olivaceus) under temperature stress.

Han P, Chen J, Sun Z, Ren S, Wang X BMC Genomics. 2025; 26(1):117.

PMID: 39920593 PMC: 11804088. DOI: 10.1186/s12864-025-11285-7.

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