诱导型一氧化氮合酶基因在虹鳟鱼（Oncorhynchus mykiss）中的表达

Expression of an inducible nitric oxide synthase gene in rainbow trout Oncorhynchus mykiss.

作者信息

Laing K J, Hardie L J, Aartsen W, Grabowski P S, Secombes C J

机构信息

Department of Zoology and Medicine & Therapeutics, University of Aberdeen, UK.

出版信息

Dev Comp Immunol. 1999 Jan-Feb;23(1):71-85. doi: 10.1016/s0145-305x(98)00036-6.

DOI:10.1016/s0145-305x(98)00036-6

PMID:10220070

Abstract

Using oligonucleotide primers based on mammalian nitric oxide synthases (NOS), expression of an inducible NOS (iNOS) gene was detected in head kidney and gill tissue of bacterially-challenged rainbow trout. Three overlapping fragments were amplified by RT-PCR and used to construct a contiguous sequence of 1410bp, with high nucleotide homology to iNOS in birds (61%) and mammals (57-59%). The nucleotide sequence translated in one reading frame to produce a partial peptide containing 470 amino acids, with 69-71% amino acid homology with mammalian iNOS, 81% homology with chicken iNOS and 85% homology with a partial (492bp) goldfish iNOS sequence. In vitro stimulation of head kidney macrophages with LPS also induced expression of the trout iNOS RNA, with optimal expression seen using 20-50 microg/ml LPS at 2h to 6h post-stimulation. The evolutionary and functional significance of the trout iNOS sequence are discussed.

摘要

使用基于哺乳动物一氧化氮合酶（NOS）的寡核苷酸引物，在受到细菌攻击的虹鳟鱼的头肾和鳃组织中检测到诱导型NOS（iNOS）基因的表达。通过逆转录聚合酶链反应（RT-PCR）扩增出三个重叠片段，并用于构建一个1410bp的连续序列，该序列与鸟类iNOS（61%）和哺乳动物iNOS（57 - 59%）具有高度核苷酸同源性。该核苷酸序列在一个阅读框中翻译，产生一个包含470个氨基酸的部分肽段，与哺乳动物iNOS的氨基酸同源性为69 - 71%，与鸡iNOS的同源性为81%，与金鱼iNOS部分序列（492bp）的同源性为85%。用脂多糖（LPS）体外刺激头肾巨噬细胞也可诱导虹鳟鱼iNOS RNA的表达，在刺激后2小时至6小时使用20 - 50μg/ml LPS时可见最佳表达。本文讨论了虹鳟鱼iNOS序列的进化和功能意义。