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Molecular characterization and expression of a crustin-like gene from Chinese mitten crab, Eriocheir sinensis
Mu, Changkao1,2; Zheng, Peilin1; Zhao, Jianmin1; Wang, Lingling1; Zhang, Huan1; Qiu, Limei1; Gai, Yunchao1; Song, Linsheng1
2010-07-01
发表期刊DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
卷号34期号:7页码:734-740
文章类型Article
摘要Crustin is a cysteine-rich antibacterial peptide which widely distributes within decapod crustaceans. In the present study, the cDNA encoding crustin-like peptide (designated CrusEs) was cloned from Chinese mitten crab Eriocheir sinensis by using rapid amplification of cDNA ends (RACE) approaches and expressed sequence tag analysis. The full-length cDNA of CrusEs was of 796 bp, containing a 5' untranslated region (UTR) of 214 bp, a 3' UTR of 267 bp with a poly(A) tail, and an open reading frame (ORF) of 315 bp encoding a polypeptide of 104 amino acids including a putative signal peptide of 21 amino acids. A WAP domain and the consensus framework existing in class I crustins were both identified in CrusEs, suggesting that CrusEs is a new member of type I crustins. Quantitative real-time RT-PCR was employed to examine the expression of CrusEs, and its mRNA transcript was mainly detected in haemocytes and gill. The CrusEs mRNA transcript in haemocytes was down-regulated after the challenge with Gram-positive bacteria Micrococcus luteus, while Gram-negative bacteria Listonella anguillarum did not induce significant variation of CrusEs mRNA. The mature peptide of CrusEs was cloned into pET-21a(+) with a C-terminal hexa-histidine tag fused in-frame, and expressed in Escherichia coli, and the recombinant CrusEs inhibited the growth of Gram-positive bacteria with low MIC. The results indicate that CrusEs is a potent antibacterial protein against Gram-positive bacteria infection, and it may play an important role in innate immune response of E. sinensis. (C) 2010 Elsevier Ltd. All rights reserved.
关键词Eriocheir Sinensis Crustin Wap Domain Recombinant Expression Antibacterial Activity Minimal Inhibitory Concentration (Mic)
DOI10.1016/j.dci.2010.02.001
收录类别SCI
语种英语
WOS记录号WOS:000277891600006
引用统计
被引频次:53[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/32793
专题实验海洋生物学重点实验室
作者单位1.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China
2.Ningbo Univ, Fac Life Sci & Biotechnol, Ningbo 315211, Zhejiang, Peoples R China
第一作者单位中国科学院海洋研究所
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Mu, Changkao,Zheng, Peilin,Zhao, Jianmin,et al. Molecular characterization and expression of a crustin-like gene from Chinese mitten crab, Eriocheir sinensis[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2010,34(7):734-740.
APA Mu, Changkao.,Zheng, Peilin.,Zhao, Jianmin.,Wang, Lingling.,Zhang, Huan.,...&Song, Linsheng.(2010).Molecular characterization and expression of a crustin-like gene from Chinese mitten crab, Eriocheir sinensis.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,34(7),734-740.
MLA Mu, Changkao,et al."Molecular characterization and expression of a crustin-like gene from Chinese mitten crab, Eriocheir sinensis".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 34.7(2010):734-740.
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