印度对虾Fein-Penaeidin的克隆、序列分析及表达

Baskaralingam Vaseeharan , Sathappan Shanthi , Jiann-Chu Chen , Montserrat Espiñeira
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引用次数: 21

摘要

对虾素是对虾体内一种特殊的抗菌肽家族成员,在对虾的免疫防御中起着重要作用。本文报道了从印度白对虾Fenneropenaus indicus (Fein-Penaeidin)中克隆的对虾蛋白序列。Fein-Penaeidin开放阅读框编码77个氨基酸的肽,包括19个氨基酸的信号肽。Fein-Penaeidin的氨基酸序列包括一个富含脯氨酸的n端结构域和一个含有6个半胱氨酸残基的羧基结构域。结构分析显示其二级结构为α -螺旋结构,预测的三维结构表明α -螺旋中存在两个二硫化物桥。系统发育分析和与其他已知花生苷的序列比较表明,该基因与单尾对虾(95%)、印度对虾(80%)和中国对虾(74%)的花生苷具有较高的相似性。Fein-Penaeidin在正常对虾和微生物攻击对虾中进行了检测,发现其在血细胞、心脏、鳃、肌肉、肠、肝胰腺和眼柄中组成性表达。细菌攻击导致mRNA上调,并在注射后6小时诱导表达,表明penaeidin参与先天免疫。
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Molecular cloning, sequence analysis and expression of Fein-Penaeidin from the haemocytes of Indian white shrimp Fenneropenaeus indicus

Penaeidins are members of a special family of antimicrobial peptide existing in penaeid shrimp and play an important role in the immunological defense of shrimp. Here, we report a penaeidin sequence cloned from the Indian white shrimp Fenneropenaus indicus (Fein-Penaeidin). The Fein-Penaeidin open reading frame encodes a 77 amino acid peptide including a 19 amino acid signal peptide. The deduced amino acid sequences of Fein-Penaeidin include a proline rich N-terminal domain and a carboxyl-domain that contains six cysteine residues. Structural analysis revealed an alpha-helix in its secondary structure and the predicted 3D structure indicated two-disulphide bridges in the alpha-helix. Phylogenetic analysis and sequence comparison with other known peaneidin suggest the gene shows high similarity to that of penaeidin from Peneaus monodon (95%), F. indicus (80%) and Fenneropenaeus chinensis (74%). Fein-Penaeidin was examined in normal and microbial challenged shrimp and was found to be constitutively expressed in haemocytes, Heart, gills, muscles, intestine, hepatopancreas and eyestalk. Bacterial challenge resulted in mRNA up-regulation, inducing expression at 6 h post injection indicating the penaeidin involved in the innate immunity.

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