军曹鱼Hepcidin基因全长cDNA克隆及其组织表达分析  

Cloning of Full Length cDNA and Tissue Expression Analysis of Hepcidin Gene from Cobia(Rachycentron canadum)

作　　者：茅莉娜[1,2] 张鹤[1] 冯娟[1] 郭志勋[1] 许海东[1] 苏友禄[1] 

机构地区：[1]中国水产科学研究院南海水产研究所农业部南海渔业资源开发利用重点实验室,广州510300 [2]广东省疾病预防控制中心,广州511430

出　　处：《中国畜牧兽医》2015年第7期1630-1639,共10页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然基金青年基金(31202027)

摘　　要：本试验采用同源克隆和末端快速扩增（RACE）的方法，得到714bp的军曹鱼（Rachycentron canadum）Hepcidin基因全长cDNA序列。该序列包括213bp的5’末端非编码区（UTR）、228bp的3’UTR及273bp的开放阅读框（ORF），编码90个氨基酸，预测其蛋白分子质量约为10．03ku，等电点为7．54，预测的蛋白包括信号肽、前肽和成熟肽。通过构建Hepcidin氨基酸序列的系统进化树并进行氨基酸同源性比对，结果显示军曹鱼与已知鱼类及哺乳动物Hepcidin氨基酸的同源性在24．4％～85．6％之间。实时荧光定量PCR（quantitative Real-timePCR，qRT-PCR）检测结果显示Hepcidin基因在正常军曹鱼组织中均表达，但表达量在各个组织中有所不同，其中以肝脏表达量最高。经脂多糖（LPS）和甲醛灭活的鲨鱼弧菌（Vibrio carchariae）刺激后，肝脏、头肾、脾脏中Hepcidi”基因表达均上调。The techniques of homology clone and RACE were used to clone the Hepcidin gene from cobia （Rachycentron canadium）. The full length cDNA of Hepcidin gene was 714 bp with a 213 bp 5runtranslated region （UTR）,a 228 bp 3PUTR and a 273 bp open reading frame （ORF） encoding a polypeptide of 90 amino acid residues with a predicted molecular weight of 10.03 ku and theoretical isoelectric point of 7.54. The predicted molecular included signal peptide, prodo- main peptide and mature peptide. Phylogenetic tree of Hepcidin amino acid sequences was con- structed and homology compapison of amino acid sequences showed that homologies were varied from 24.4% to 85. 6% with some known Hepcidin amino acids in other fishes and mammals. Quantitative Real-time PCR （qRT-PCR） analysis revealed that Hepcidin gene was expressed in all tissues with different expression levels,which expressed most in liver. The Hepcidin gene ex-pressions in liver,head kidney and spleen were up-regulated after stimulation of LPS and forma- lin-inactivated Vibrio carchariae.

关 键 词：军曹鱼 HEPCIDIN基因 全长CDNA 组织表达 

分 类 号：Q78[生物学—分子生物学]