Key Laboratory of Mariculture (Ministry of Education), Ocean University of China, Qingdao 266003, Shandong, PR China.
Comp Biochem Physiol C Toxicol Pharmacol. 2010 Nov;152(4):424-32. doi: 10.1016/j.cbpc.2010.07.002. Epub 2010 Jul 18.
A novel cDNA encoding ferritin (HdhNFT) was cloned from the hepatopancreas of abalone, Haliotis discus hannai Ino. The deduced protein contains 171 amino acid residues with a predicted molecular mass (MW) about 19.8 kDa and theoretical isoelectric point (pI) of 4.792. Amino acid alignment revealed that HdhNFT shared high similarity with other known ferritins. The HdhNFT contained a highly conserved motif for the ferroxidase center, which consists of seven residues of a typical vertebrate heavy-chain ferritin with a typical stem-loop structure. HdhNFT mRNA contains a 27 bp iron-responsive element (IRE) in the 5'-untranslated region. This IRE exhibited 82.14% similarity with abalone H. discus discus and 78.57% similarity with Pacific oyster Crassostrea gigas IREs. By real-time PCR assays, the mRNA transcripts of HdhNFT were found to be higher expressed in kidney, hepatopancreas, gill, mantle and muscle than in haemocytes and gonad. Moreover, mRNA expression levels of HdhNFT in the hepatopancreas and haemocytes were measured by real-time PCR in abalone fed with graded levels of dietary iron (29.2, 65.7, 1267.2 and 6264.7 mg/kg). Results showed that the expression of the HdhNFT mRNA increased with dietary iron contents. Furthermore, the maximum value of the HdhNFT mRNA was found in the treatment with 6264.7 mg/kg of dietary iron. These data indicated that dietary iron can up-regulate HdhNFT at transcriptional level in abalone.
从鲍鱼(Haliotis discus hannai Ino)的肝胰腺中克隆了一种编码铁蛋白(HdhNFT)的新 cDNA。推导的蛋白质包含 171 个氨基酸残基,预测的分子量(MW)约为 19.8 kDa,理论等电点(pI)为 4.792。氨基酸比对表明,HdhNFT 与其他已知铁蛋白具有高度相似性。HdhNFT 包含一个高度保守的铁氧化酶中心基序,由典型脊椎动物重链铁蛋白的七个残基组成,具有典型的茎环结构。HdhNFT mRNA 在 5'非翻译区含有 27 个碱基的铁反应元件(IRE)。该 IRE 与鲍鱼 H. discus discus 的相似度为 82.14%,与太平洋牡蛎 Crassostrea gigas 的 IRE 的相似度为 78.57%。通过实时 PCR 分析,发现 HdhNFT 的 mRNA 转录物在肾脏、肝胰腺、鳃、套膜和肌肉中的表达高于血淋巴细胞和性腺。此外,通过实时 PCR 测量了鲍鱼在不同水平的膳食铁(29.2、65.7、1267.2 和 6264.7 mg/kg)下肝胰腺和血淋巴细胞中 HdhNFT 的 mRNA 表达水平。结果表明,HdhNFT mRNA 的表达随膳食铁含量的增加而增加。此外,在膳食铁含量为 6264.7 mg/kg 的处理中,HdhNFT mRNA 的最大值。这些数据表明,膳食铁可以在转录水平上调鲍鱼中的 HdhNFT。