Cloning and recombinant expression of a crustin-like gene from Chinese shrimp, Fenneropenaeus chinensis
Cloning and recombinant expression of a crustin-like gene from Chinese shrimp, Fenneropenaeus chinensis
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DOI：
10.1016/j.jbiotec.2006.08.013
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发表时间：
2007-01-20
影响因子：
4.1
通讯作者：
Xiang, Jianhai
中科院分区：
文献类型：
--
作者：
Zhang, Jiquan;Li, Fuhua;Xiang, Jianhai
抗菌肽或抗菌蛋白（AMPs）已被证明是抵抗病原体感染最重要的体液因子之一。作为一种抗菌蛋白，壳蛋白在无脊椎动物中被描述为先天免疫系统的一个组成部分。采用3′和5′-RACE PCR技术，从中国对虾（Fenneropenaeus chinensis）的血细胞中克隆了一个壳蛋白样基因（CruFc）。全长cDNA由523个长度为405 bp的开放阅读框组成，编码134个氨基酸，推断出的肽含有17个氨基酸的推定信号肽。该序列在c端还含有一个乳清酸性蛋白（WAP）结构域。通过RT-PCR分析，主要在血细胞和鳃中检测到CruFc转录本。此外，从中国对虾的血细胞中克隆了另一个全长cDNA CshFc，其推断的氨基酸序列缺乏wap型“四二硫核”结构域。分别制备了含有CruFc和CshFc的融合蛋白，抑菌实验结果表明，在相同条件下，重组CruFc能抑制谷物阳性菌的体外生长，而重组CshFc不能抑制。重组CruFc和CshFc的抑菌活性差异表明，壳蛋白的四二硫核结构域可能在其生物学功能中发挥重要作用。(c) 2006 Elsevier B.V.版权所有
Antimicrobial peptides or proteins (AMPs) are proved to be one of the most important humoral factors to resist pathogen infection. As an antimicrobial protein, crustin had been described in invertebrates as a component of the innate immune system. A crustin-like gene (CruFc) was cloned from haemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5'-RACE PCR. The full-length cDNA consists of 523 with 405 bp open reading frame encoding 134 amino acids and the deduced peptide contains a putative signal peptide of 17 amino acids. The sequence also contains a whey-acidic protein (WAP) domain at the C-terminal. Transcripts of CruFc were mainly detected in haemocytes and gill by RT-PCR analysis. In addition, another full-length cDNA named CshFc was also cloned from haemocytes of Chinese shrimp and its inferred amino acid sequence lacks the WAP-type 'four-disulfide core' domain. The fusion proteins containing CruFc and CshFc were, respectively, produced and the antimicrobial assays revealed that the recombinant CruFc could inhibit the growth of grain-positive bacteria in vitro but the recombinant CshFc could not inhibit at the same conditions. The difference of antimicrobial activity between recombinant CruFc and CshFc provides the evidence that the four-disulfide core domain of crustin may play an important role in its biological function. (c) 2006 Elsevier B.V. All rights reserved.