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果糖-1,6-二磷酸酶(fructose-1,6-bisphosphatase,FBP,EC 3.1.3.11)可催化果糖-1,6-二磷酸水解成果糖-6-磷酸和无机磷酸盐,是糖异生途径中的关键酶之一。本研究运用SMART RACE技术从鲈鱼Lateolabrax japonicus肝脏中分离克隆了FBP基因的全长cDNA序列,该基因全长1 357bp,其中5’非翻译区和3’非翻译区分别为42bp和301bp,开放阅读框为1 014bp,共编码337个氨基酸。蛋白质分子量约为36.7kD,理论pI为6.90。氨基酸序列分析表明,鲈鱼FBP与其它动物的肝脏型FBP相似性很高,与裸盖鱼、彩虹胡瓜鱼、斑马鱼、异育银鲫和大西洋鲑的肝脏型FBP的同源性分别为94.3%,90.8%,89.3%,88.1%和84.1%。系统发育分析显示,鲈鱼FBP与其它鱼类的肝脏型FBP成簇后再与哺乳动物的肝脏型FBP聚成一支,然后才与哺乳动物的肌肉型FBP汇成簇。同时用RT-PCR分析了FBP基因在鲈鱼肝脏、肌肉、心脏、眼、肠、肾脏、脂肪、脾脏、鳃和大脑等10个组织的表达,结果表明FBP仅在肝脏、肾脏和肠这3个组织中有较高的表达,与糖异生发生组织基本一致,因此推测该FBP属于肝脏型。
Fructose-1,6-bisphosphatase (FBP, EC 3.1.3.11) catalyzes the hydrolysis of fructose-1,6-diphosphate to fructose-6-phosphate and inorganic phosphates, One of the key enzymes in the biological pathway. In this study, the full-length cDNA of FBP gene was isolated and purified from the liver of Lateolabrax japonicus by SMART RACE. The full-length cDNA of FBP gene is 1 357bp in length, including 42bp and 301bp in 5 ’untranslated region and 3’ untranslated region, respectively. The frame is 1 014 bp, encoding a total of 337 amino acids. The protein has a molecular weight of about 36.7 kD and a theoretical pI of 6.90. Amino acid sequence analysis showed that the sera of FBP in sea bass were highly similar to those of other animals, and the homologies with FBP in liver of naked bonito, rainbow smelt, zebrafish, allogynogenetic crucian carp and Atlantic salmon were 94.3% , 90.8%, 89.3%, 88.1% and 84.1%. Phylogenetic analysis showed that the sea bass FBP was clustered with liver fibrils of other fishes and clustered together with mammalian liver FBPs, before clustering with mammalian muscle-type FBPs. The expression of FBP gene in ten tissues of liver, muscle, heart, eye, intestine, kidney, fat, spleen, gill and brain of sea bass was analyzed by RT-PCR. The results showed that FBP was only expressed in liver, kidney and intestine Higher expression in the tissue, and gluconeogenesis is basically the same, so speculated that the FBP belongs to the liver type.