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日本对虾中新的无脊椎动物溶菌酶的鉴定与表达分析

Identification and expression analysis of a new invertebrate lysozyme in Kuruma shrimp (Marsupenaeus japonicus).

作者信息

Liu Hong-Tao, Wang Jun, Mao Yong, Liu Min, Niu Su-Fang, Qiao Ying, Su Yong-Quan, Wang Chun-Zhong, Zheng Zhi-Peng

机构信息

State Key Laboratory of Marine Environmental Sciences, Xiamen University, Xiamen, Fujian 361102, China.

State Key Laboratory of Marine Environmental Sciences, Xiamen University, Xiamen, Fujian 361102, China.

出版信息

Fish Shellfish Immunol. 2016 Feb;49:336-43. doi: 10.1016/j.fsi.2015.12.034. Epub 2015 Dec 24.

Abstract

Lysozyme is an important component of the innate immunity system against invading pathogens. An invertebrate (i-type) lysozyme from the hepatopancreas of Kuruma shrimp Marsupenaeus japonicus (Mj-ilys) was identified. The full-length cDNA of Mj-ilys was 580bp with a 429 bp open reading frame encoding a 142 amino acid polypeptide. The encoded polypeptide was predicted to have a 17 amino acid signal peptide, and a 125 amino acid mature protein with a theoretical mass of 14.099 kDa and an isoelectric point (pI) of 4.18. A Destabilase conserved domain was predicted in Mj-ilys amino acid sequences which may be stable by 10 cysteine residues forming 5 disulfide bonds. Mj-ilys may loss the muramidase and isopeptidase activities due to the lack of the key catalytic residues. Mj-ilys had high homologous of 80-82% with i-type lysozymes of penaeid shrimps. It was first grouped with other i-type lysozyme of shrimps and crabs in a phylogenetic tree predicted by the Neighbor-Joining method. Mj-ilys mRNA was expressed mainly in hepatopancreas and almost undetectable in other tissues. The mRNA expression of Mj-ilys were all found from fertilized eggs to post-larvae of 17 days (PL17), and its expression exhibited significant differences among each developmental stage. After white spot syndrome virus (WSSV) challenge (3.6 × 10(8) virions/μl), the time-dependent expression pattern of Mj-ilys in hepatopancreas and gills showed significantly different. These results indicated that Mj-ilys is potentially involved in the ontogenesis and immune defense in Kuruma shrimp.

摘要

溶菌酶是针对入侵病原体的天然免疫系统的重要组成部分。已从日本对虾(Marsupenaeus japonicus)的肝胰腺中鉴定出一种无脊椎动物(i型)溶菌酶(Mj-ilys)。Mj-ilys的全长cDNA为580bp,具有429bp的开放阅读框,编码一个142个氨基酸的多肽。预测该编码多肽具有一个17个氨基酸的信号肽和一个125个氨基酸的成熟蛋白,理论分子量为14.099kDa,等电点(pI)为4.18。在Mj-ilys氨基酸序列中预测到一个Destabilase保守结构域,其可能通过10个半胱氨酸残基形成5个二硫键而保持稳定。由于缺乏关键催化残基,Mj-ilys可能丧失溶菌酶和异肽酶活性。Mj-ilys与对虾i型溶菌酶具有80-82%的高度同源性。在通过邻接法预测的系统发育树中,它首先与虾和蟹的其他i型溶菌酶归为一组。Mj-ilys mRNA主要在肝胰腺中表达,在其他组织中几乎检测不到。从受精卵到17日龄后期幼体(PL17)均能检测到Mj-ilys的mRNA表达,且其在各发育阶段的表达存在显著差异。在感染白斑综合征病毒(WSSV)(3.6×10(8)个病毒粒子/μl)后,Mj-ilys在肝胰腺和鳃中的时间依赖性表达模式显示出显著差异。这些结果表明,Mj-ilys可能参与了日本对虾的个体发育和免疫防御。

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