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首页> 外文期刊>Fish & Shellfish Immunology >Hsp60 in caudal fin regeneration from Paramisgurnus dabryanus: Molecular cloning and expression characterization
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Hsp60 in caudal fin regeneration from Paramisgurnus dabryanus: Molecular cloning and expression characterization

机译：HSP60在尾鳍尾鳍再生中的分子克隆和表达表征

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摘要

Heat shock protein 60 (Hsp60) is a kind of highly conserved immunogenic molecule involved in a wide range of biochemical processes in response to external stressors. Its multifunction in regulating immune responses and modulating signal pathway interests us in investigating its role in fin regeneration that has become an excellent and interesting model for studying the molecular basis of morphogenesis. We firstly clarified basical process and crucial period of caudal fins regeneration in Paramisgurnus dabryanus by histological analysis. Then we cloned full-length cDNA of hsp60 from P. dabryanus (designated as PdHsp60) by RACE method. The cDNA contains a 124 bp 5'UTR, a 1731 bp open reading frame (ORF) encoding 576 amino acids and a 510bp 3'UTR (Accession no.: KF544774). The phylogenetic tree shows that the PdHsp60 fits within the hsp60 clade. And quantitative RT-PCR detected the PdHsp60 began to increase rapidly its expression at 1 dpa and reached its peak at 2 dpa. Next, spatial distribution analysis of PdHsp60 in fins showed that PdHsp60 located mainly in the deeper lay of regenerated epidermis when PdHsp60 expressed most. After the PdHsp60 had been cloned into the pET-32a vector, SDS-PAGE and Western blotting analysis confirmed that the PdHsp60 protein was efficiently expressed in Escherichia coli BL21. These findings have revealed that PdHsp60, a highly conserved gene related to the innate immune system and stress response during vertebrate evolution, is involved in response to wounding stimulation-in the formation of wound epidermis which occurs as the first phase of fin regeneration after fin amputation in caudal fin regeneration.
机译：热休克蛋白60（Hsp60）是一种高度保守的免疫原性分子，涉及多种生物化学过程，以应对外界应激。它在调节免疫应答和调节信号途径中的多功能性使我们感兴趣，以研究其在鳍再生中的作用，该鳍已成为研究形态发生的分子基础的极好且有趣的模型。首先，通过组织学分析，弄清了黄Para尾鳍再生的基本过程和关键时期。然后，我们通过RACE方法从黄杨假单胞菌中克隆了hsp60的全长cDNA（命名为PdHsp60）。该cDNA含有124bp的5'UTR，1731bp的开放阅读框（ORF），其编码576个氨基酸和510bp的3'UTR（登录号：KF544774）。系统发育树表明，PdHsp60适合在hsp60进化枝中。定量RT-PCR检测到PdHsp60在1 dpa时开始迅速增加表达，在2 dpa时达到峰值。其次，鳍中PdHsp60的空间分布分析表明，当PdHsp60表达最多时，PdHsp60主要位于再生表皮的较深层。将PdHsp60克隆到pET-32a载体后，SDS-PAGE和Western印迹分析证实PdHsp60蛋白在大肠杆菌BL21中有效表达。这些发现表明，PdHsp60是一种高度保守的基因，与脊椎动物进化过程中的先天免疫系统和应激反应有关，它参与了对伤口刺激的反应-在伤口表皮的形成中，这是在鳍截肢后鳍再生的第一阶段。在尾鳍再生中。

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