该研究通过同源克隆技术，成功克隆并测序了海鳕（Melanogrammus aeglefinus）的两种促炎细胞因子——IL-1β和IL-8的基因。海鳕IL-1β的全长转录本含有1043个碱基对，包含一个762个碱基对的开放阅读框。IL-8的cDNA包含903个碱基对，其预测蛋白质序列在所有已知的硬骨鱼基因中首次显示出ELRmotif。这两种细胞因子在不同组织（头肾、脾脏、肝脏和鳃）中的表达受到刺激的影响，IL-1β主要由PMA和LPS026:B6诱导表达，而IL-8则显示出恒定表达，但表达水平受刺激物质调控。
摘要：
This paper reports the cloning and sequencing of two pro-inflammatory cytokines, interleukin (IL)-1beta and IL-8, in haddock (Melanogrammus aeglefinus) by homology cloning. The complete transcript of the haddock IL-1beta was sequenced and contained 1043 bp, including a 762 bp open reading frame. The 3' end of the gene includes a polyadenylation signal 13 bp upstream of the poly(A) tail, along with 10 instability motifs. The predicted protein of 253aa revealed the presence of the IL-1 family signature and the absence of an ICE cut site. The cDNA of the chemokine IL-8 was sequenced in haddock and contained 903 bp of which 306 bp are the open reading frame. Interestingly, the predicted protein sequence of 101aa, contains an ELR motif preceding the CXC signature, common in all vertebrate IL-8 molecules but absent in all teleost genes sequenced to date. The expression of both haddock cytokines was studied in four different tissues: head kidney, spleen, liver and gill. Tissues were obtained from both healthy fish and fish stimulated in vivo with four commercial serotypes of LPS, namely Escherichia coli 026:B6, 055:B5, 0111:B4 and 0127:B8 and PMA. Haddock IL-1beta was not constitutively expressed and expression was only observed following stimulation. However, this expression was stimulant dependent and only PMA and LPS 026:B6 induced high levels of expression in the head kidney. The haddock IL-8 gene on the other hand, showed a constitutive expression, that could be up or down-regulated depending on the immunostimulant used, although to a lesser extent than IL-1beta.
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