Identification and molecular characterization of Cathepsin L gene and its expression analysis during early ontogenetic development of kuruma shrimp Marsupenaeus japonicus
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摘要: 本研究从日本囊对虾(Marsupenaeus japonica)中获得半胱氨酸蛋白酶家族中的cathepsin L基因,命名为Mj-Cathepsin L。Mj-Cathepsin L基因cDNA序列全长1191bp,其开放阅读框编码的Cathepsin L蛋白前体包含327个氨基酸,由信号肽,蛋白前体和成熟蛋白三部分组成。同源序列比对发现日本囊对虾Cathepsin L蛋白与其他已知物种的cahtepsin L蛋白具有较高的同源性。对Mj-Cathepsin L基因在不同组织以及早期幼体幼体发育阶段的研究发现,Mj-Cathepsin L主要在肝胰腺中大量表达,且在幼体发育不同阶段,其表达水平受发育阶段调控,且Mj-Cathepsin L基因显示出与其五个蜕皮时期相关的表达变化,说明Mj-Cathepsin L有可能在日本囊对虾个体发育过程中扮演不同的重要作用。并通过Western Blot方法在日本囊对虾肝胰腺,胃和肠中检测到Cathepsin L酶原和成熟Cathepsin L两种蛋白存在形式。
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关键词:
- 日本囊对虾 /
- Mj-CathepsinL /
- 幼体发育 /
- 蜕皮阶段 /
- 组织表达 /
- Marsupenaeusjaponicus
Abstract: Cathepsin L gene is a member of the cysteine proteinase gene group. In this study Cathepsin L gene was isolated from Kuruma shrimp Marsupenaeus japonicus (Mj-Cathepsin L) and the full-length DNA sequence was 1 963 bp. Mj-Cathepsin L protein showed high homologies with other Cathepsin L proteins documented in vertebrates, mollusks and other crustaceans. Expression analysis of Mj-Cathepsin L gene in different tissues revealed that it was predominant in hepatopancreas. During early ontogenetic development stages Mj-Cathepsin L showed a development-regulated expression, and the Mj-Cathepsin L showed a molting stage-regulated expression during the five molting stages, inferring its role in the ontogenic development of M. japonicus. Two kinds of forms of Mj-Cathepsin L protein:pro-Cathepsin L and Cathepsin L were measured in hepatopancreas, stomach and intestine by Western Blotting.-
Key words:
- Cathepsin L gene /
- larval development /
- molting cycle /
- tissue distribution /
- Marsupenaeus japonicus /
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