摘要
采用同源克隆和锚定PCR技术,从军曹鱼(RachycentroncanadiumLinnaeus)中克隆到白细胞介素1β(interleukin 1β)基因cDNA的全序列。军曹鱼IL 1β的cDNA全长1104bp,3′非编码区域(UTR)为255bp,5′UTR为108bp,开放阅读框(ORF)为741bp,编码246个氨基酸,分子量大约为27 68kD,理论等电点为5 71。同源性分析表明,该序列与其他鱼类甚至哺乳动物的IL 1β基因具有很高的相似性,并含有白细胞介素家族的签名序列(signature)。同时,利用RT PCR技术,对特异性病原刺激下该基因在鱼体内的表达情况进行初步研究,发现IL 1β基因在鱼体的多种组织中都有表达,而经过脂多糖(lipopolysaccharide,LPS)刺激后,目的基因在组织中的表达明显增加,但是在不同组织内的表达存在差异。研究显示,军曹鱼IL 1β基因存在组成型和诱导型2种表达调控机制,在抵御病原感染过程中发挥重要作用。
IL-1β is a pleiotropic proinlammatory cytokine with a wide spectrum of inflammatory,metabolic,hematopoietic and immunological activities and is responsible for the symptoms of sickness during the host response to infection.IL-1β is produced by many cell types,including monocytes,macrophages,langerhans cells,dendritic cells,endothelial,epithelial cells and fibroblasts,even in sperm.Tremendous progress has been made in gene cloning of cytokines from fish in recent years,especially TNFα and IL-1β et al.Several evidences provided by biological cross reaction have strongly suggested that interleukins exist in fish.IL-1β bioactivity has been known in fish for over a decade,but only since 1999,IL-1β gene has been cloned in a number of teleost species and at the same time confirmed those initial findings.To date most analysis has been made on rainbow trout(Oncorhynchus mykiss) and carp(Cyprinus carpio) genes.More recently,seabass(Dicentrarchus labrax),seabream(Sparus aurata),turbot(Scophthalmus maximus),dogfish(Scyliorhinus caniculus) and catshark(Scyliorhinus canicula) IL-1β genes have been cloned and sequenced.The techniques of homology cloning and anchored PCR were used to clone the IL-1β gene from cobia(Rachycentron canadium Linnaeus).The full length cDNA of IL-1β is 1104bp,containing a 5' untranslated region (UTR) of 108bp,an ORF of 741bp,a encoding polypeptide of 246 amino acids with an estimated molecular mass of 27.68kD,and a 3' UTR of 255bp.The searches for nucleotides and protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of cobia IL-1β was homologic to the IL-1β in other fish species and even the mammalian.Conserved signature sequences of IL-1β gene family and several potential glycosylation sites were found in the cobia IL-1β deduced amino acid sequence.Analysis with the Signal P software revealed that there was no signal peptide in the sequence,which was common with the other known IL-1β molecules.Just as other nonmammalian IL-1β genes sequenced to
出处
《中国水产科学》
CAS
CSCD
北大核心
2005年第2期119-125,共7页
Journal of Fishery Sciences of China
基金
广东省科技攻关项目(2003C20313)
农业部海洋与河口渔业重点开放实验室开放基金项目
广东省农业攻关项目(2003B21502).
