Leibniz-Institut für Nutztierbiologie (FBN), Fachbereich Molekularbiologie, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany.
Mol Biol Rep. 2012 Apr;39(4):4291-300. doi: 10.1007/s11033-011-1216-1. Epub 2011 Jul 22.
The Ca(2+)-binding protein regucalcin (RGN) is crucial for the regulation of Ca(2+) ion homeostasis and signal transduction of cells. It is involved in the regulation of Ca(2+)-dependent protein kinases and Ca(2+) pump enzymes in cell membranes. Comparative transcriptome analysis in healthy fish of two aquacultured rainbow trout (Oncorhynchus mykiss) lines (BORN, TCO) varying in susceptibility to environmental stress identified significant differences in the expression of the RGN gene. Therefore, we firstly determined the full genomic DNA and cDNA sequence of RGN gene from rainbow trout and comparatively investigated the complete cDNA sequence in another salmonid fish dedicated for local aquaculture, the maraena whitefish (Coregonus marena). The sequence coding region translates for proteins of 298 and 299 amino acids (aa), respectively, indicating a high conservation of RGN proteins (95.7% aa identity) between the two related salmonids. In the second place, we generated RGN gene expression profiles after pathogen (Aeromonas salmonicidae subsp. salmonicida) and temperature (8 and 23°C) challenge in the two rainbow trout lines using salmon microarrays and quantitative RT-PCR. The profiles not only verified initially detected gene expression differences, they also display a tissue specific gene expression in dependence from the stressor and time. The differences in gene expression support our assumption that RGN might play a role in recovery of rainbow trout after environmental stress.
钙结合蛋白 Regucalcin（RGN）对于细胞内钙离子稳态和信号转导的调节至关重要。它参与调节细胞膜中钙离子依赖性蛋白激酶和钙离子泵酶。对两种对环境胁迫敏感性不同的养殖虹鳟（Oncorhynchus mykiss）品系（BORN、TCO）的健康鱼进行比较转录组分析，发现 RGN 基因的表达存在显著差异。因此，我们首先从虹鳟中确定了 RGN 基因的全长基因组 DNA 和 cDNA 序列，并比较研究了另一种专门用于当地水产养殖的鲑鱼——白鲑（Coregonus marena）中的完整 cDNA 序列。编码区序列分别翻译为 298 和 299 个氨基酸（aa）的蛋白质，表明两种相关鲑鱼之间的 RGN 蛋白高度保守（95.7% aa 同一性）。其次，我们使用鲑鱼微阵列和定量 RT-PCR 在两种虹鳟品系中，在病原体（鲑鱼气单胞菌亚种）和温度（8 和 23°C）挑战后，生成了 RGN 基因表达谱。这些图谱不仅验证了最初检测到的基因表达差异，还显示了组织特异性基因表达，这取决于应激源和时间。基因表达的差异支持我们的假设，即 RGN 可能在虹鳟从环境胁迫中恢复后发挥作用。
