摘要
精氨酸酶II(arginase II,Arg-II)是精氨酸酶的一种,不仅可以参与机体尿素循环,还与机体病理过程密切相关。通过香鱼(Plecoglossus altivelis)单核/巨噬细胞转录组测序获得香鱼Arg-II基因全长c DNA序列,结果表明,Arg-II基因由4 707个核苷酸组成,包含一个大的开放阅读框,编码348个氨基酸,预测分子量为38.09 k D,等电点为6.15。氨基酸序列多重比对结果显示,香鱼Arg-Ⅱ具有典型的Arg-Ⅱ结构特征,与虹鳟(Oncorhynchus mykiss)Arg-Ⅱ同源性最高,为85.3%;系统进化树分析表明,鱼类Arg-Ⅱ形成一个大簇,香鱼Arg-Ⅱ与虹鳟Arg-Ⅱ进行相关性最高。实时荧光定量PCR(quantitive real-time PCR,q RTPCR)结果显示,香鱼Arg-Ⅱ基因m RNA主要在健康香鱼肝、脑、头肾和单核/巨噬细胞中表达;鳗弧菌(Vibrio anguillarum)感染后,香鱼肝、脑、头肾和单核/巨噬细胞Arg-Ⅱ基因m RNA的表达量显著上调。综上,香鱼Arg-Ⅱ基因的表达与鳗弧菌感染紧密相关,揭示其可能在鱼类抗感染免疫应答中发挥重要作用。
Arginase II（Arg-II）is one kind of arginases. Not only can it participates in the urea cycle, but also has close correlation with pathological process. Here, we determined the whole c DNA sequence of Arg-II gene of ayu（Plecoglossus altivelis）by de-novo transcriptome sequencing of ayu monocytes/macrophages. The results showed that the c DNA sequence of the Arg-II was 4707 nucleotides, containing a large open reading frame that encoded a protein of 348 amino acids with a deduced molecular mass of 38.09 k D and a theoretical isoelectric point（p I）of 6.15. Multiple sequence alignment of complete amino acid sequences showed that ayu Arg-II had the typical characters of Arg-II in animals. Sequence comparison showed that ayu Arg-II shared the highest amino acid sequence identity（85.3%）with that of rainbow trout（Oncorhynchus mykiss）. Phylogenetic tree analysis also confirmed that fish Arg-II formed a cluster, and ayu Arg-II was most closely related to that of rainbow trout. Quantitative real-time PCR（q RT-PCR）analysis showed that the m RNA of ayu Arg-II mainly had the expression in liver, brain, head kidney and monocytes/macrophages. Upon Vibrio anguillarum infection, the m RNA expression of gene Arg-II significantly up-regulated in the liver, brain, head kidney and monocytes/macrophages. In summary, our present study revealed a tight relationship between the ayu Arg-II expression and V. anguillarum infection, suggesting that ayu Arg-II plays an important role in fish immune response against bacterial infection. This study provides a theoretical basis for studying the functions of fish Arg-II, and its molecular mechanism in regulating fish immune response to pathogens.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第2期109-115,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(31402333)
浙江省自然科学基金项目(LQ13C190002)
高等学校博士学科点专项科研基金项目(20133305120008)
关键词
香鱼
精氨酸酶II
鳗弧菌
单核/巨噬细胞
表达
Plecoglossus altivelis
arginase II
Vibrio anguillarum
monocytes/macrophage
expression
