To understand the role of 14-3-3 beta/alpha in hypoosmotic regulation of turbot (Scophthalmus maximus), we characterized the 14-3-3 beta/alpha gene and analyzed the tissue distribution and its gene transcriptional patterns in the main expressed tissues under low salt stress. The 14-3-3 beta/alpha cDNA is 892 bp in length, incorporating an ORF of 774 bp with a putative primary structure of 257 residues. The deduced amino acid sequences shared highly conserved structures with other eukaryotes. Quantitative real-time PCR results showed that the 14-3-3 beta/alpha transcripts were widely expressed in various tissues of turbot, with most abundant in the gill (P < .05), to a lesser extent in the kidney, intestine, brain and spleen, and at low levels in the pituitary and other tissues examined. And the expression of turbot 14-3-3 beta/alpha exhibited a trend of increasing first and then decreasing with the time of stress under low salt stress, and the highest value appeared in 12 h (P < .05). After injecting different concentrations of dsRNA, the mRNA expression of 14-3-3 gene decreased significantly during the monitoring period, and the best interference effect was achieved 12 h after injecting 4 μg/g dsRNA. For the first time, the gene was silenced in fish by intramuscular injection of dsRNA. It also provides a new and effective way to study gene function at the individual level. Moreover, the mRNA interference of 14-3-3 beta/alpha would cause changes in the expression of several ion channel proteins, for example, the decrease of Na+-K+-ATPase and Na+-H+-exchanger and the increase of CFTR. As a result, 14-3-3 beta/alpha appears to be an important molecular regulator for osmosensory signal transduction in gill of turbot.

中文翻译：

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大菱t（Scophthalmus maximus）的分子表征，14-3-3 beta / alpha的表达分析以及RNA干扰对离子转运蛋白Na + -K + -ATPase，Na + -H +-交换子和CFTR的影响。

为了了解14-3-3 beta / alpha在低通透性比目鱼（Scophthalmus maximus）中的作用，我们对14-3-3 beta / alpha基因进行了表征，并分析了主要表达组织中的组织分布及其基因转录模式在低盐胁迫下。14-3-3 beta / alpha cDNA的长度为892 bp，并整合了774 bp的ORF和一个假定的257个残基的一级结构。推导的氨基酸序列与其他真核生物具有高度保守的结构。实时定量PCR结果显示14-3-3 beta / alpha转录本在大菱turbo的各种组织中广泛表达，其中abundant最丰富（P <.05），而在肾脏，肠、,中的表达较少。脑和脾脏，并在垂体和其他组织中低水平检查。低盐胁迫下，大菱1414-3-3 beta / alpha的表达呈现出先升高后降低的趋势，并在12 h达到最大值（P <.05）。注入不同浓度的dsRNA后，在监测期间14-3-3基因的mRNA表达显着下降，并且在注入4μg/ g dsRNA后12 h达到了最佳的干扰效果。首次通过肌肉内注射dsRNA沉默了该基因。它还为研究个体水平的基因功能提供了一种新的有效途径。此外，mRNA干扰14-3-3 beta / alpha会引起几种离子通道蛋白表达的变化，例如，Na + -K + -ATPase和Na + -H +-交换子的减少以及CFTR的增加。结果是，