Hu Rong, Qu Fufa, Tang Jianzhou, Zhao Qiong, Yan Jinpeng, Zhou Zhigang, Zhou Yi, Liu Zhen
Department of Biological and Environmental Engineering, Changsha University, Changsha, Hunan 410022, China.
Department of Cell Biology, School of Life Sciences, Central South University, Changsha 410017, China.
Comp Biochem Physiol B Biochem Mol Biol. 2017 Oct;212:70-76. doi: 10.1016/j.cbpb.2017.06.004. Epub 2017 Jun 19.
Glutamine synthetase (GS) is considered a master enzyme that catalyzes ATP-dependent biosynthesis of glutamine from glutamate. In the present study, the GS gene was cloned from the intestine of grass carp (Ctenopharyngodon idellus). The full-length cDNA sequence of GS encodes a 371-amino-acid polypetide. Phylogenetic analysis of the C. idellus GS sequence reveals common carp (Cyprinus carpio) as its closest neighbor. GS mRNA was differentially expressed in different tissues, with high to low gradient expression the intestine, brain, muscle, heart, gill, liver, pituitary gland, and spleen. Additionally, GS exhibited a dynamic pattern of expression during embryonic development, reaching maximal and minimal levels in the organ and hatching stages, respectively, and constant low levels from 7 to 28days post-hatching. We also assessed dietary protein levels and feed sources in diet-regulated fish, and the results suggested that low crude protein (CP) and fish meal stimulate GS gene expression. Furthermore, intestinal GS mRNA expression was significantly increased by 0.2, 0.4, 0.6, 0.8mM concentrations of glutamine dipeptide in vitro. This study provides valuable knowledge about the regulation of GS expression in teleosts.
谷氨酰胺合成酶(GS)被认为是一种催化由谷氨酸通过ATP依赖性生物合成谷氨酰胺的关键酶。在本研究中,GS基因是从草鱼(Ctenopharyngodon idellus)的肠道中克隆出来的。GS的全长cDNA序列编码一个371个氨基酸的多肽。对草鱼GS序列的系统发育分析表明,鲤鱼(Cyprinus carpio)是其最接近的亲缘物种。GS mRNA在不同组织中差异表达,在肠道、脑、肌肉、心脏、鳃、肝脏、垂体和脾脏中呈从高到低的梯度表达。此外,GS在胚胎发育过程中呈现出动态表达模式,分别在器官形成期和孵化期达到最高和最低水平,在孵化后7至28天保持低水平。我们还评估了饲料调控鱼的日粮蛋白质水平和饲料来源,结果表明低粗蛋白(CP)和鱼粉会刺激GS基因表达。此外,在体外,0.2、0.4、0.6、0.8mM浓度的谷氨酰胺二肽可显著提高肠道GS mRNA的表达。本研究为硬骨鱼中GS表达的调控提供了有价值的知识。