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从软体动物长牡蛎中克隆和表达两种肽聚糖识别蛋白(PGRP)基因。

Molecular cloning and mRNA expression of two peptidoglycan recognition protein (PGRP) genes from mollusk Solen grandis.

机构信息

Shandong Marine Fisheries Research Institute, Yantai 264006, China.

出版信息

Fish Shellfish Immunol. 2012 Jan;32(1):178-85. doi: 10.1016/j.fsi.2011.11.009. Epub 2011 Nov 20.

Abstract

Peptidoglycan recognition proteins (PGRPs) play crucial role in innate immunity for both invertebrates and vertebrates, owing to their prominent ability in detecting and eliminating invading bacteria. In the present study, two short PGRPs from mollusk Solen grandis (designated as SgPGRP-S1 and SgPGRP-S2) were identified, and their expression patterns, both in tissues and toward three PAMPs stimulation, were then characterized. The full-length cDNA of SgPGRP-S1 and SgPGRP-S2 was 1672 and 1285 bp, containing an open reading frame (ORF) of 813 and 426 bp, respectively, and deduced amino acid sequences showed high similarity to other members of PGRP superfamily. Both SgPGRP-S1 and SgPGRP-S2 encoded a PGRP domain. The motif of Zn(2+) binding sites and amidase catalytic sites were well conserved in SgPGRP-S1, but partially conserved in SgPGRP-S2. The two PGRPs exhibited different tissue expression pattern. SgPGRP-S1 was highly expressed in muscle and hepatopancreas, while SgPGRP-S2 was highly in gill and mantle. The mRNA expression of SgPGRP-S1 could be induced acutely by stimulation of PGN, and also moderately by β-1,3-glucan, but not by LPS, while expression of SgPGRP-S2 was significantly up-regulated (P < 0.01) when S. grandis was stimulated by all the three PAMPs, though the expression levels were relatively lower than SgPGRP-S1. Our results suggested SgPGRP-S1 and SgPGRP-S2 could serve as pattern recognition receptors (PRRs) involved in the immune recognition of S. grandis, and they might perform different functions in the immune defense against invaders.

摘要

肽聚糖识别蛋白(PGRPs)在无脊椎动物和脊椎动物的先天免疫中发挥着至关重要的作用,这要归功于它们在检测和消除入侵细菌方面的突出能力。在本研究中,从软体动物大竹蛏(Solen grandis)中鉴定出两种短肽聚糖识别蛋白(PGRPs)(分别命名为 SgPGRP-S1 和 SgPGRP-S2),并对其在组织中的表达模式以及对三种 PAMP 刺激的反应进行了表征。SgPGRP-S1 和 SgPGRP-S2 的全长 cDNA 分别为 1672bp 和 1285bp,分别含有 813bp 和 426bp 的开放阅读框(ORF),推导的氨基酸序列与 PGRP 超家族的其他成员高度相似。SgPGRP-S1 和 SgPGRP-S2 均编码一个 PGRP 结构域。SgPGRP-S1 中的 Zn(2+)结合位点和酰胺酶催化位点的基序保守较好,而 SgPGRP-S2 中的部分保守。这两种 PGRPs 表现出不同的组织表达模式。SgPGRP-S1 在肌肉和肝胰腺中高表达,而 SgPGRP-S2 在鳃和套膜中高表达。PGN 刺激可使 SgPGRP-S1 的 mRNA 表达迅速诱导,β-1,3-葡聚糖也可适度诱导,但 LPS 则不能诱导,而 S. grandis 受到这三种 PAMP 刺激时,SgPGRP-S2 的表达显著上调(P < 0.01),尽管其表达水平相对低于 SgPGRP-S1。我们的研究结果表明,SgPGRP-S1 和 SgPGRP-S2 可作为参与大竹蛏免疫识别的模式识别受体(PRRs),它们在抵抗入侵物的免疫防御中可能发挥不同的功能。

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