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碱性磷酸酶(alkaline phosphatase,ALP)是一种广泛分布的单脂磷酸水解酶,是动物生物代谢过程中重要的调控酶。实验首次克隆了大黄鱼ALP基因cDNA全序列,命名为LcALP,其全长为2 345 bp,开放阅读框为1 629 bp,编码543个氨基酸。实时荧光定量PCR检测发现,LcALP在大黄鱼雌雄各组织中均有表达,其中在眼和头肾的表达量最高,雌性鳃和脑中LcALP表达量显著高于雄性,而在性腺和脾脏中的表达量则显著低于雄性;胚胎发育过程中,多细胞期、囊胚期和原肠期LcALP基因表达水平相对较低,在卵黄栓形成期明显提高,至孵出期达到峰值,而初孵仔鱼期则显著下调;大黄鱼肌肉细胞系经LPS处理后6 h LcALP表达量降低,12 h显著下调,而poly I:C处理后表达水平持续上升,24 h显著上调至峰值。研究表明,LcALP在大黄鱼胚胎发育调控以及机体免疫防御中发挥重要作用。
Alkaline phosphatase (ALP) is a widely distributed monolipid hydrolase, which is an important regulatory enzyme in animal biological metabolism. The full length cDNA of ALP gene of large yellow croaker was cloned for the first time and named LcALP. The full length cDNA was 2 345 bp, with an open reading frame of 1 629 bp encoding 543 amino acids. Real-time PCR showed that LcALP was expressed in both male and female tissues of large yellow croaker, with the highest expression in eye and head kidney, and the expression of LcALP in gill and brain of female was significantly higher than that in male, while in gonad and spleen The expression level of LcALP gene in multi-cell stage, blastula stage and gastrula stage was relatively lower during embryogenesis, significantly increased in the stage of yolk thrombosis and peaked at hatching stage, while the initial hatch The expression of LcALP was decreased 6 h after LPS treatment in large yellow croaker muscle cells and significantly down-regulated at 12 h. However, the expression of LcALP increased continuously after the treatment of poly I: C, and reached a peak at 24 h. Studies have shown that LcALP plays an important role in the regulation of embryonic development and immune defense in large yellow croaker.