香鱼巨噬细胞炎性蛋白-2基因的克隆、序列分析及表达  

Molecular Cloning,Sequence Analysis and Prokaryotic Expression of Sweetfish Macrophage Inflammatory Protein-2 Gene

作　　者：王世会[1] 史雨红[1] 陈炯[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,宁波315211

出　　处：《生物技术通报》2014年第7期143-149,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31372555)

摘　　要：巨噬细胞炎性蛋白-2(MIP-2)是一种重要的趋化因子,可趋化中性粒细胞到炎症部位,从而消除炎症反应。从香鱼巨噬细胞转录组测序中获得MIP-2基因,阅读框序列长为318个核苷酸,编码一个由105个氨基酸组成、相对分子质量为11.6kD的前体蛋白。N端19个氨基酸为信号肽序列。氨基酸序列分析表明,香鱼MIP-2与白斑狗鱼MIP-2的氨基酸同源性最高,为54%。健康香鱼MIP-2基因mRNA主要在脾、肾、脑、鳃中表达,在肝、心、肌肉、肠中表达量次之。实时荧光定量PCR结果显示,鳗利斯顿氏菌侵染香鱼后,各组织中MIP-2基因mRNA表达量均呈上调趋势。尤其以肾组织和肌肉组织中变化最显著。以上结果表明,香鱼MIP-2基因表达与鳗利斯顿氏菌的侵染密切相关,揭示了MIP-2可能在香鱼抗菌免疫反应中具有重要的作用。Macrophage inflammatory protein-2（MIP-2）is an important chemokine, which promotes neutrophils to be the sites of inflammation and eliminates the inflammatory response. In this study, the sequence of MIP-2 gene from de-novo transcriptome sequencing of sweetfish macrophages was obtained. An open reading frame was composed of 318 nucleotides, which encoded a protein of 105 amino acids with a molecular weight of 11.6 kD. Its N-terminal 19 residues were the signal peptides. Sequence alignment showed that sweetfish MIP-2 shared the highest homology with the MIP-2 from Esox lucius with 54%protein sequence identity. In healthy sweetfish, MIP-2 mRNA was mainly expressed in the spleen, kidney, brain and gills, weakly expressed in the liver, heart, muscle and intestine. Quantitative RT-PCR analysis showed that MIP-2 transcripts were significantly up-regulated in the tested tissues, especially the kidney and muscle. The results suggested that MIP-2 might play an important role in sweetfish antibacterial immunity.

关 键 词：巨噬细胞炎性蛋白-2香鱼 鳗利斯顿氏菌感染 基因表达 

分 类 号：S917.4[农业科学—水产科学]