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Molecular cloning and characterization of a serine proteinase homolog prophenoloxidase-activating factor in the swimming crab Portunus trituberculatus

文献类型：期刊论文

作者Cui, Zhaoxia1; Liu, Yuan1,2; Wu, Danhua1,3; Luan, Weisha1,2; Wang, Shuangyan1,2; Li, Qianqian1,2; Song, Chengwen1,2
刊名FISH & SHELLFISH IMMUNOLOGY
出版日期2010-10-01
卷号29期号:4页码:679-686
关键词Portunus Trituberculatus Serine Protein Homologous Eyestalk Haemocyte
ISSN号1050-4648
DOI10.1016/j.fsi.2010.07.015
文献子类Article
英文摘要Serine proteinase homologues (SPHs), as one of prophenoloxiase-activating factors (PPAFs), play critical roles in innate immunity of crabs. Based on an EST from the eyestalk full length cDNA library, the complete cDNA (designated as PtSPH) and genomic DNA of SPH from the swimming crab Portunus trituberculatus were cloned in this study. The estimated molecular weight of mature PtSPH (354 amino acids) was 38.7 kDa and its isoelectric point was 5.08. Multiple sequence alignment revealed that PtSPH lacked a catalytic residue with a substitution of Ser in the active site triad to Gly. Phylogenetic analysis indicated PtSPH together with PPAFs of Callinectes sapidus (AAS60227), Eriocheir sinensis (ACU65942), Penaeus monodon (ABE03741, ACP19563) and Pacifastacus leniusculus (ACB41380), formed a distinct cluster which only included clip-SPHs. As the first analyzed genomic structure of PPAFs in crustaceans, two introns were found in the open reading frame region of this gene. The mRNA transcripts of PtSPH could be detected in all the examined tissues, and were higher expressed in the eyestalk than that in gill, hepatopancreas, haemocytes and muscle. Accompanied with the change in phenoloxidase (PO) activity and total haemocyte counts, the temporal expression of PtSPH gene in haemocytes after Vibrio alginolyticus challenge demonstrated a clear time-dependent expression pattern with two peaks within the experimental period of 32 h. These findings suggest that PtSPH is involved in the antibacterial defense mechanism of Portunus tritubercualtus crab. (C) 2010 Elsevier Ltd. All rights reserved.; Serine proteinase homologues (SPHs), as one of prophenoloxiase-activating factors (PPAFs), play critical roles in innate immunity of crabs. Based on an EST from the eyestalk full length cDNA library, the complete cDNA (designated as PtSPH) and genomic DNA of SPH from the swimming crab Portunus trituberculatus were cloned in this study. The estimated molecular weight of mature PtSPH (354 amino acids) was 38.7 kDa and its isoelectric point was 5.08. Multiple sequence alignment revealed that PtSPH lacked a catalytic residue with a substitution of Ser in the active site triad to Gly. Phylogenetic analysis indicated PtSPH together with PPAFs of Callinectes sapidus (AAS60227), Eriocheir sinensis (ACU65942), Penaeus monodon (ABE03741, ACP19563) and Pacifastacus leniusculus (ACB41380), formed a distinct cluster which only included clip-SPHs. As the first analyzed genomic structure of PPAFs in crustaceans, two introns were found in the open reading frame region of this gene. The mRNA transcripts of PtSPH could be detected in all the examined tissues, and were higher expressed in the eyestalk than that in gill, hepatopancreas, haemocytes and muscle. Accompanied with the change in phenoloxidase (PO) activity and total haemocyte counts, the temporal expression of PtSPH gene in haemocytes after Vibrio alginolyticus challenge demonstrated a clear time-dependent expression pattern with two peaks within the experimental period of 32 h. These findings suggest that PtSPH is involved in the antibacterial defense mechanism of Portunus tritubercualtus crab. (C) 2010 Elsevier Ltd. All rights reserved.
学科主题Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
URL标识查看原文
语种英语
WOS记录号WOS:000281500500018
公开日期2010-12-24
源URL[http://ir.qdio.ac.cn/handle/337002/5879]  
专题海洋研究所_海洋生物技术研发中心
海洋研究所_实验海洋生物学重点实验室
作者单位1.Chinese Acad Sci, Inst Oceanol, EMBL, Qingdao 266071, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China
3.Ningbo Univ, Ningbo 315000, Zhejiang, Peoples R China
推荐引用方式
GB/T 7714		 Cui, Zhaoxia,Liu, Yuan,Wu, Danhua,et al. Molecular cloning and characterization of a serine proteinase homolog prophenoloxidase-activating factor in the swimming crab Portunus trituberculatus[J]. FISH & SHELLFISH IMMUNOLOGY,2010,29(4):679-686.
APA Cui, Zhaoxia.,Liu, Yuan.,Wu, Danhua.,Luan, Weisha.,Wang, Shuangyan.,...&Song, Chengwen.(2010).Molecular cloning and characterization of a serine proteinase homolog prophenoloxidase-activating factor in the swimming crab Portunus trituberculatus.FISH & SHELLFISH IMMUNOLOGY,29(4),679-686.
MLA Cui, Zhaoxia,et al."Molecular cloning and characterization of a serine proteinase homolog prophenoloxidase-activating factor in the swimming crab Portunus trituberculatus".FISH & SHELLFISH IMMUNOLOGY 29.4(2010):679-686.

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来源：海洋研究所

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