Characterization and expression of sweetfish （Plecoglossus altivelis） cathepsin D

Characterization and expression of sweetfish （Plecoglossus altivelis） cathepsin D

Cathepsin D （CTSD） is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish （Plecoglossus altivelis） CTSD （PaCTSD） via de-novo transcriptome sequencing of sweetfish macrophages. The full length cDNA sequence of PaCTSD was 1955 bp enco...

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Veröffentlicht in:动物学研究 2014, Vol.35 (4), p.294-299
1. Verfasser: Yu JIAO Chang-Hong LI Xin-Jiang LU Jiong CHEN
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Sprache:chi
Schlagworte:
免疫应答
全长cDNA序列
实时定量PCR
氨基酸序列
系统进化树分析
组织蛋白酶D
表征
香鱼
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description Cathepsin D （CTSD） is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish （Plecoglossus altivelis） CTSD （PaCTSD） via de-novo transcriptome sequencing of sweetfish macrophages. The full length cDNA sequence of PaCTSD was 1955 bp encoding a propeptide of 397 amino acids. The deduced protein had a calculated molecular weight of 43.17x 103. Multiple alignment with other known CTSD amino acid sequences revealed amino acid conservation through the teleosts. Phylogenetic tree analysis showed that PaCTSD grouped tightly with other fish CTSD, and was close to that of Atlantic salmon and rainbow trout. Subsequently, PaCTSD was prokaryotically expressed and refolded by the urea gradient method on a nickel-nitrilotriacetic acid column. Enzyme activity analysis showed that PaCTSD exhibited pH-dependent proteolytic activity. Quantitative real-time PCR showed that PaCTSD mRNA was expressed in all detected tissues in healthy sweetfish. The highest expression
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The full length cDNA sequence of PaCTSD was 1955 bp encoding a propeptide of 397 amino acids. The deduced protein had a calculated molecular weight of 43.17x 103. Multiple alignment with other known CTSD amino acid sequences revealed amino acid conservation through the teleosts. Phylogenetic tree analysis showed that PaCTSD grouped tightly with other fish CTSD, and was close to that of Atlantic salmon and rainbow trout. Subsequently, PaCTSD was prokaryotically expressed and refolded by the urea gradient method on a nickel-nitrilotriacetic acid column. Enzyme activity analysis showed that PaCTSD exhibited pH-dependent proteolytic activity. Quantitative real-time PCR showed that PaCTSD mRNA was expressed in all detected tissues in healthy sweetfish. 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source Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Bioline International; PubMed Central
subjects 免疫应答
全长cDNA序列
实时定量PCR
氨基酸序列
系统进化树分析
组织蛋白酶D
表征
香鱼
title Characterization and expression of sweetfish （Plecoglossus altivelis） cathepsin D
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