College of Life Sciences, Liaoning Normal University, 1, Liushu South Street, Ganjingzi District, Dalian, 116081, People's Republic of China.
Mol Biol Rep. 2014 Jan;41(1):9-18. doi: 10.1007/s11033-013-2764-3. Epub 2013 Sep 20.
Glycogen serves as a metabolic reserve and is involved in macromolecular synthesis. Glycogen phosphorylase (GPase) is a key enzyme involved in intracellular glycogen catabolism, catalyzing the first step in glycogen degradation. In the diapause, GPase catalyzes glycogen into the closely related molecule, sorbitol. In this study, the full-length cDNA of the GPase gene (2,790 bp) was isolated from Artemia sinica for the first time by rapid amplification of cDNA ends technology. The GPase gene encoded a protein of 853 amino acids belonging to the Glycosyltransferase GTB type superfamily. The expression pattern and location of GPase were investigated at various stages during the embryonic development of A. sinica using real-time PCR and in situ hybridization. High GPase expression was detected at the 0 and 5 h stages. Subsequently, expression declined and was maintained at a low level during the stages from 10 to 40 h following by a small increase at day 3. Expression was downregulated at temperatures ranging from 25 to 20 °C and was subsequently upregulated in the range 15-5 °C. In situ hybridization assays showed wide distribution of the GPase gene during different developmental stages. From the results of this study, we conclude that the GPase gene expression is stress-related and might play an important role in Artemia development and metabolism.
糖原作为代谢储备物质，参与大分子的合成。糖原磷酸化酶（GPase）是参与细胞内糖原分解代谢的关键酶，催化糖原降解的第一步。在休眠期，GPase 将糖原催化成与之密切相关的分子，山梨醇。在这项研究中，通过快速扩增 cDNA 末端技术，首次从中华卤虫中分离出全长的 GPase 基因（2790bp）。GPase 基因编码的蛋白含有 853 个氨基酸，属于糖基转移酶 GTB 超家族。使用实时 PCR 和原位杂交技术研究了 GPase 在中华卤虫胚胎发育的各个阶段的表达模式和位置。在 0 和 5h 阶段检测到高 GPase 表达。随后，表达水平下降并在 10-40h 阶段维持在低水平，随后在第 3 天略有增加。在 25-20°C 的温度范围内表达下调，随后在 15-5°C 的范围内上调。原位杂交分析显示 GPase 基因在不同发育阶段广泛表达。根据本研究的结果，我们得出结论，GPase 基因的表达与应激有关，可能在卤虫的发育和代谢中发挥重要作用。
