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草鱼(Ctenopharyngodon idella)中寡肽转运蛋白(PepT1)的特性研究。

Characterization of oligopeptide transporter (PepT1) in grass carp (Ctenopharyngodon idella).

机构信息

Department of Biotechnology and Environmental Science, Changsha University, Changsha 410003, China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2013 Mar;164(3):194-200. doi: 10.1016/j.cbpb.2012.11.008. Epub 2012 Dec 4.

Abstract

The oligopeptide transporter (PepT1) is located on the brush-border membrane of the intestinal epithelium, and plays an important role in dipeptide and tripeptide absorptions from protein digestion. In this study, we cloned the PepT1 cDNA from grass carp and characterized its expression profile in response to dietary protein and feed additives (sodium butyrate) treatments. The PepT1 gene encodes a protein of 714 amino acids with high sequence similarity with other vertebrate homologues. Expression analysis revealed highest levels of PepT1 mRNA expression in the foregut of grass carp. In addition, PepT1 mRNA expression exhibited diurnal variation in all three bowel segments of intestine with lower levels of expression in daytime than nighttime. During embryonic development, PepT1 showed a dynamic pattern of expression reaching maximal levels of expression in the gastrula stage and minimal levels in the organ stage. The PepT1 expression showed constant levels from 14 to 34 day post-hatch. To determine whether fish diet of different protein contents may have any effect on PepT1 expression, we extended our research to dietary regulation of PepT1 expression. We found that dietary protein levels had a significant effect on PepT1 gene expression. In addition, PepT1 mRNA levels were higher after feeding with fish meal than with soybean meal. Moreover, in vitro and in vivo sodium butyrate treatments increased PepT1 expression in the intestine of grass carp. The results demonstrate for the first time that PepT1 mRNA expression is regulated in a temporal and spatial pattern during development, and dietary protein and feed additives had a significant effects on PepT1 gene expression in grass carp.

摘要

寡肽转运蛋白(PepT1)位于肠上皮细胞的刷状缘膜上,在二肽和三肽从蛋白质消化中吸收方面发挥着重要作用。在这项研究中,我们从草鱼中克隆了 PepT1 cDNA,并研究了其在日粮蛋白和饲料添加剂(丁酸钠)处理下的表达模式。PepT1 基因编码一个 714 个氨基酸的蛋白质,与其他脊椎动物同源物具有高度的序列相似性。表达分析表明,PepT1 mRNA 表达水平在草鱼的前肠最高。此外,PepT1 mRNA 表达在所有三个肠段都具有昼夜变化,白天的表达水平低于夜间。在胚胎发育过程中,PepT1 表现出动态表达模式,在原肠胚阶段达到最高表达水平,在器官阶段达到最低表达水平。从孵化后 14 天到 34 天,PepT1 的表达水平保持恒定。为了确定不同蛋白含量的鱼类日粮是否会对 PepT1 表达产生影响,我们将研究扩展到日粮对 PepT1 表达的调控。我们发现,日粮蛋白水平对 PepT1 基因表达有显著影响。此外,与大豆蛋白相比,鱼粉喂养后 PepT1 mRNA 水平更高。此外,体外和体内丁酸钠处理增加了草鱼肠道中 PepT1 的表达。结果首次证明,PepT1 mRNA 表达在发育过程中具有时空模式的调节,并且日粮蛋白和饲料添加剂对草鱼 PepT1 基因表达有显著影响。

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