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王英慧, 修云吉, 顾伟, 孟庆国, 王文. 日本沼虾两种抗脂多糖因子的特性研究[J]. 水生生物学报, 2017, 41(5): 977-983. DOI: 10.7541/2017.122
引用本文: 王英慧, 修云吉, 顾伟, 孟庆国, 王文. 日本沼虾两种抗脂多糖因子的特性研究[J]. 水生生物学报, 2017, 41(5): 977-983. DOI: 10.7541/2017.122
WANG Ying-Hui, XIU Yun-Ji, GU Wei, MENG Qing-Guo, WANG Wen. MOLECULAR CLONING, CHARACTERIZATION, AND EXPRESSION ANALYSIS OF TWO ISOFORMS OF ANTI-LIPOPOLYSACCHARIDE FACTOR FROM THE ORIENTAL RIVER PRAWN, MACROBRACHIUM NIPPONENSE[J]. ACTA HYDROBIOLOGICA SINICA, 2017, 41(5): 977-983. DOI: 10.7541/2017.122
Citation: WANG Ying-Hui, XIU Yun-Ji, GU Wei, MENG Qing-Guo, WANG Wen. MOLECULAR CLONING, CHARACTERIZATION, AND EXPRESSION ANALYSIS OF TWO ISOFORMS OF ANTI-LIPOPOLYSACCHARIDE FACTOR FROM THE ORIENTAL RIVER PRAWN, MACROBRACHIUM NIPPONENSE[J]. ACTA HYDROBIOLOGICA SINICA, 2017, 41(5): 977-983. DOI: 10.7541/2017.122

日本沼虾两种抗脂多糖因子的特性研究

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

  • 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MnALF1、MnALF2。MnALF1 cDNA 全长1008 bp, 编码121个氨基酸; MnALF2 cDNA 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MnALFs与之前发现的甲壳动物的ALFs是非常相似的。qRT-PCR结果显示MnALFs在所有被检测的组织中均有表达。其中MnALF1主要在心脏和小肠内表达, 而MnALF2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MnALFs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(MnALF1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MnALF2, 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MnALF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。

     

    Abstract: Anti-lipopolysaccharide factors (ALFs), a type of the potent antimicrobial peptide, can bind and neutralize lipopolysaccharide (LPS) and exhibit broad spectrum antimicrobial activities. In order to study the function of ALFs in congenital immunization of Macrobrachium nipponense, two isoforms of the ALF homologues (MnALF1 and MnALF2) were cloned and characterized from the oriental river prawn M. nipponense. The full-length cDNA sequences of MnALF1 and MnALF2 were 1008 and 836 bp, encoding 121 and 124 amino acids, respectively. All of these sequences contained one signal peptide and an LPS-binding domain with two conserved cysteine residues at both ends of the domain. The deduced peptide of MnALF1 and MnALF2 was highly similar to previously identified ALFs in crustaceans. qRT-PCR showed that MnALFs were expressed in all detected tissues. MnALF1 transcript was predominantly detected in heart and intestine and MnALF2 transcript was predominantly detected in hemocytes and hepatopancreas. After challenge with Aeromonas hydrophila, two MnALF transcripts in heart, intestine, hemocytes and hepatopancreas showed a clear time-dependent response expression pattern (the expression levels of MnALF1 present an trend of down-regulated first and then increasing, which reached to the highest level at 24h, 12h, 36h, 24h in heart, intestine, hemocytes and hepatopancreas respectively. However, for MnALF2, the expression level present an down-regulated trend and then increased in heart, intestine; In hemocytes and hepatopancreas, the expression level present an increasing trend and then down-regulated. MnALF2 transcripts reached to the top at 24h post-challenge). These results suggest that two MnALF isoforms have different tissue specificity and might provide multiple protective functions in immune defense against invading bacteria.

     

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