日本沼虾两种抗脂多糖因子的特性研究（英文）  

MOLECULAR CLONING, CHARACTERIZATION, AND EXPRESSION ANALYSIS OF TWO ISOFORMS OF ANTI-LIPOPOLYSACCHARIDE FACTOR FROM THE ORIENTAL RIVER PRAWN, MACROBRACHIUM NIPPONENSE

作　　者：王英慧[1] 修云吉 顾伟[1] 孟庆国[1] 王文[1] WANG Ying-Hui;XIU Yun-Ji;GU Wei;MENG Qing-Guo;WANG Wen(Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases,College of Life Sciences, Nanjing Normal University, Nanjing 210023, China;Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences,Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China)

机构地区：[1]南京师范大学生命科学学院,江苏省生物多样性与生物技术重点实验室,江苏省水生甲壳动物病害重点实验室,南京210023 [2]中国水产科学研究院黄海水产研究所,农业部海洋渔业可持续发展重点实验室,青岛266071 [3]海洋科学与技术国家实验室,海洋渔业科学与食物产出过程功能实验室,青岛266200

出　　处：《水生生物学报》2017年第5期977-983,共7页Acta Hydrobiologica Sinica

基　　金：Supported by the National Natural Science Foundation of China(31570176,31602198);the Natural Science Foundation of Jiangsu Province(BK20151545);Project for Aquaculture in Jiangsu Province(D2015-13,Y2016-28)

摘　　要：为了研究抗脂多糖因子ALFs在日本沼虾先天性免疫中的功能作用,研究从日本沼虾中克隆了2种抗脂多糖因子Mn ALF1、Mn ALF2。Mn ALF1 c DNA全长1008 bp,编码121个氨基酸;Mn ALF2 c DNA全长836 bp,编码124个氨基酸。这2种氨基酸均包含有一个信号肽序列和一个LPS结合位点,并且在结合位点的两端(N-端和C-端)都有2个保守的半胱氨酸残基。这2种Mn ALFs与之前发现的甲壳动物的ALFs是非常相似的。q RT-PCR结果显示Mn ALFs在所有被检测的组织中均有表达。其中Mn ALF1主要在心脏和小肠内表达,而Mn ALF2则主要在血细胞和肝胰脏中表达。在用嗜水气单胞菌刺激之后发现2种Mn ALFs在心脏、小肠、血细胞、肝胰脏中都呈现出明显的时间依赖表达模式(Mn ALF1在刺激之后呈现出先减少后增加的趋势,之后分别在不同组织的不同时间点达到最大值;然而,对于Mn ALF2,在心脏和小肠中先减少后增加,在血细胞和肝胰脏中呈现出先增加后减少,最后都在24h达到最大值)。结果提示这2种Mn ALF具有不同的组织特异性,并且在细菌侵染的免疫防御中起着重要的保护作用。Anti-lipopolysaccharide factors(ALFs),a type of the potent antimicrobial peptide,can bind andneutralize lipopolysaccharide(LPS)and exhibit broad spectrum antimicrobial activities.In order to study thefunction of ALFs in congenital immunization of Macrobrachium nipponense,two isoforms of the ALFhomologues(MnALF1and MnALF2)were cloned and characterized from the oriental river prawn M.nipponense.The full-length cDNA sequences of MnALF1and MnALF2were1008and836bp,encoding121and124amino acids,respectively.All of these sequences contained one signal peptide and an LPS-binding domainwith two conserved cysteine residues at both ends of the domain.The deduced peptide of MnALF1andMnALF2was highly similar to previously identified ALFs in crustaceans.qRT-PCR showed that MnALFswere expressed in all detected tissues.MnALF1transcript was predominantly detected in heart and intestineand MnALF2transcript was predominantly detected in hemocytes and hepatopancreas.After challenge withAeromonas hydrophila,two MnALF transcripts in heart,intestine,hemocytes and hepatopancreas showed aclear time-dependent response expression pattern(the expression levels of MnALF1present an trend of downregulatedfirst and then increasing,which reached to the highest level at24h,12h,36h,24h in heart,intestine,hemocytes and hepatopancreas respectively.However,for MnALF2,the expression level present an downregulatedtrend and then increased in heart,intestine;In hemocytes and hepatopancreas,the expression levelpresent an increasing trend and then down-regulated.MnALF2transcripts reached to the top at24h post-challenge).These results suggest that two MnALF isoforms have different tissue specificity and might providemultiple protective functions in immune defense against invading bacteria.

关 键 词：日本沼虾 抗脂多糖因子 表达分析 嗜水气单胞菌 

分 类 号：Q344.1[生物学—遗传学]