Identification of housekeeping genes suitable for gene expression analysis in the pearl mussel, Hyriopsis cumingii, during biomineralization
Identification of housekeeping genes suitable for gene expression analysis in the pearl mussel, Hyriopsis cumingii, during biomineralization
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DOI：
10.1007/s00438-014-0837-1
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发表时间：
2014-08-01
影响因子：
3.1
通讯作者：
Li, Jiale
中科院分区：
文献类型：
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作者：
Bai, Zhiyi;Lin, Jingyun;Li, Jiale
实时定量聚合酶链反应是一种灵敏的定量基因表达水平的技术。必须选择一个或多个适当的参考基因，以准确地比较不同样品和组织中的mRNA转录水平。三角帆蚌（Hyriopsis cumingii，莱亚）是我国重要的养殖经济品种。迄今为止，尚未验证用于该物种基因表达分析的参考基因。本研究旨在比较7个管家基因在不同组织类型和在外套膜或珍珠囊中的相对表达，在三个生物矿化过程中：季节性贝壳生长，贝壳愈合和珍珠囊形成。cumingii。三个程序评估了七个基因的表达稳定性：BestKeeper、geNorm和Normandy。β肌动蛋白基因（ACTB）在许多研究中通常用作管家基因，是最不稳定的。泛素（Ubi）、核糖体蛋白L18（Rpl 18）和延伸因子1-α（EF 1 α）的表达比其余四个基因更稳定。因此，我们建议Ubi、Rpl 18和EF 1 α是合适的参考基因。这三个参考基因有望为分析珍珠母贝形成过程中的基因表达提供参考。cumingii。
Quantitative real-time polymerase chain reaction is a sensitive technique for quantifying gene expression levels. One or more appropriate reference genes must be selected to accurately compare mRNA transcript levels across different samples and tissues. The freshwater pearl, Hyriopsis cumingii (Lea), is an important economic species cultured in China. To date, no reference genes for gene expression analysis in this species have been validated. This study aimed to compare the relative expression of seven housekeeping genes across different tissue types and in the mantle or pearl sac during three biomineralization processes: seasonal shell growth, shell healing and pearl-sac formation in H. cumingii. Three programs evaluated the expression stabilities of the seven genes: BestKeeper, geNorm and NormFinder. The beta actin gene (ACTB), commonly used as a housekeeping gene in many studies, was the least stable. The expressions of Ubiquitin (Ubi) and Ribosomal protein L18 (Rpl18) and Elongation factor 1-alpha (EF1 alpha) were more stable than the remaining four genes. Therefore, we suggest that Ubi, Rpl18 and EF1 alpha are suitable reference genes. The three selected reference genes are expected to facilitate analysis of gene expressions during shell or pearl formation in H. cumingii.