School of Veterinary Science, The University of Queensland, Gatton, QLD, Australia.
School of Animal and Veterinary Sciences, The University of Adelaide, Roseworthy, SA, Australia.
Sci Rep. 2018 Feb 20;8(1):3364. doi: 10.1038/s41598-018-21723-0.
To better understand host and immune response to diseases, gene expression studies require identification of reference genes with stable expression for accurate normalisation. This study describes the identification and testing of reference genes with stable expression profiles in koala lymph node tissues across two genetically distinct koala populations. From the 25 most stable genes identified in transcriptome analysis, 11 genes were selected for verification using reverse transcription quantitative PCR, in addition to the commonly used ACTB and GAPDH genes. The expression data were analysed using stable genes statistical software - geNorm, BestKeeper, NormFinder, the comparative ΔCt method and RefFinder. All 13 genes showed relative stability in expression in koala lymph node tissues, however Tmem97 and Hmg20a were identified as the most stable genes across the two koala populations.
为了更好地了解宿主和免疫对疾病的反应，基因表达研究需要鉴定具有稳定表达谱的参考基因，以进行准确的归一化。本研究描述了在两个遗传上不同的考拉种群的考拉淋巴结组织中，具有稳定表达谱的参考基因的鉴定和测试。在转录组分析中确定的 25 个最稳定的基因中，选择了 11 个基因进行逆转录定量 PCR 验证，除了常用的 ACTB 和 GAPDH 基因。使用稳定基因统计软件 - geNorm、BestKeeper、NormFinder、比较 ΔCt 方法和 RefFinder 分析表达数据。所有 13 个基因在考拉淋巴结组织中的表达均表现出相对稳定性，但 Tmem97 和 Hmg20a 被鉴定为两个考拉种群中最稳定的基因。
