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Reference gene selection for transcriptional profiling in Cryptocercus punctulatus, an evolutionary link between
Zhen Li1,2, Xiangrui Li2,3, Qingwen Zhang1
1Department of Entomology and MOA Key Lab of Pest Monitoring and Green Management, China Agricultural University, Beijing, China.
Scientific Reports
|December 18, 2020
Summary
This study identifies ACT as the most stable reference gene for gene expression analysis in the wood-feeding cockroach, Cryptocercus punctulatus. This finding is crucial for accurate genetic studies on cockroach evolution and lignocellulase acquisition.
Area of Science:
- Evolutionary biology
- Genomics
- Molecular biology
Background:
- Cryptocercus cockroaches offer insights into the evolution of eusociality and wood-feeding.
- Accurate gene expression quantification requires reliable normalization strategies.
- RT-qPCR is a key technique for genetic analysis.
Purpose of the Study:
- To identify stable reference genes for RT-qPCR in Cryptocercus punctulatus.
- To establish a standardized method for gene expression analysis in this species.
- To support genetic research on cockroach evolution and lignocellulase acquisition.
Main Methods:
- Selection of 10 candidate housekeeping genes from the Cryptocercus gut transcriptome.
- Analysis of gene expression profiles using geNorm, NormFinder, BestKeeper, and comparative ΔCT methods.
- Comprehensive ranking of reference gene stability using RefFinder.
Main Results:
- ACT was identified as the most stable reference gene across different developmental stages and tissues.
- The stability of reference genes significantly impacted the analysis of target genes (Hex-1 and Cell-1).
Conclusions:
- ACT is a reliable reference gene for RT-qPCR in Cryptocercus.
- Standardized RT-qPCR protocols are essential for accurate genetic research in Cryptocercus.
- This study provides a foundational step for future genetic investigations in this species.
