用于鲤鱼和金鱼多倍体基因表达分析的最优参考基因。
Optimal reference genes for gene expression analysis in polyploid of Cyprinus carpio and Carassius auratus.
机构信息
State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, P.R. China.
College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, P.R. China.
出版信息
BMC Genet. 2020 Sep 17;21(1):107. doi: 10.1186/s12863-020-00915-6.
BACKGROUND
Reference genes are usually stably expressed in various cells and tissues. However, it was reported that the expression of some reference genes may be distinct in different species. In this study, we intend to answer whether the expression of reported traditional reference genes changes or not in the polyploid fish RESULTS: By retrieving the mRNA sequencing data of three different ploidy fish from the NCBI SRA database, we selected 12 candidate reference genes, and examined their expression levels in the 10 tissues and in the four cell lines of three different ploidy fish by real-time PCR. Then, the expression profiles of these 12 candidate reference genes were systematically evaluated by using the software platforms: BestKeeper, NormFinder and geNorm.
CONCLUSION
The 28S ribosomal protein S5 gene (RPS5) and the ribosomal protein S18 gene (RPS18) are the most suitable reference genes for the polyploid of Cyprinus carpio and Carassius auratus, demonstrated by both of the tissues and the cultured cells.
背景
参考基因通常在各种细胞和组织中稳定表达。然而，据报道，一些参考基因的表达在不同物种中可能存在差异。在本研究中，我们旨在回答报告的传统参考基因的表达是否在多倍体鱼类中发生变化。
结果
通过从 NCBI SRA 数据库中检索三种不同倍性鱼类的 mRNA 测序数据，我们选择了 12 个候选参考基因，并通过实时 PCR 检测了它们在三种不同倍性鱼类的 10 种组织和 4 种细胞系中的表达水平。然后，使用软件平台 BestKeeper、NormFinder 和 geNorm 系统地评估了这 12 个候选参考基因的表达谱。
结论
28S 核糖体蛋白 S5 基因（RPS5）和核糖体蛋白 S18 基因（RPS18）是鲤鱼和金鱼多倍体最适合的参考基因，这在组织和培养细胞中均得到了验证。
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