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首页> 外文期刊>African Journal of Biotechnology >Molecular cloning, characterization and expression analysis of heat shock protein 90 (HSP90) from the mud crab Scylla paramamosain
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Molecular cloning, characterization and expression analysis of heat shock protein 90 (HSP90) from the mud crab Scylla paramamosain

机译：泥蟹Sylla paramamosain热休克蛋白90（HSP90）的分子克隆，表征和表达分析

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Heat shock protein 90 (HSP90) is a highly conserved protein and plays an important role in maintaining the structure of protein, participating in the immunity and regulating the cell cycle. Using the rapid amplification of cDNA ends (RACE) techniques, the cDNA sequence of HSP90 gene (designated?Sp-HSP90) was cloned and characterized from the mud crab?Scylla paramamosain. The full-length cDNA of?Sp-HSP90?is 2677 bp with a complete open reading frame (ORF) of 2166 bp, which encodes a polypeptide of 721 amino acids. Five conserved blocks defining HSP90 protein family were found in the deduced amino acid sequence of?Sp-HSP90.?It contains an adenosine-5’-triphosphatase (ATPase) domain in the N-terminal and a conserved signature sequence MEEVD in the C-terminal. Quantitative real-time polymerase chain reaction (PCR) (qRT-PCR) analyses revealed the distribution of?Sp-HSP90?mRNA in different tissues and its temporal expression in haemocytes of the crabs challenged with?Vibrio parahaemolyticus. Different levels of?Sp-HSP90?mRNA were detected in heart, hepatopancreas, muscle, haemocytes, testis and ovary except eyestalk. The expression level of?Sp-Hsp90?mRNA in hemocytes was found to be obviously up-regulated after live and heat-killed bacterial challenge and significantly higher in live bacteria group than that in heat-killed bacteria group. These results suggest that?Sp-HSP90?gene might act on immunity and resistance to infection in?S. paramamosain.
机译：热休克蛋白90（HSP90）是高度保守的蛋白，在维持蛋白结构，参与免疫力和调节细胞周期中起着重要作用。利用cDNA末端快速扩增（RACE）技术，从泥蟹？Sylyl paramamosain克隆并鉴定了HSP90基因的cDNA序列（命名为？Sp-HSP90）。 Sp-HSP90β的全长cDNA为2677bp，具有2166bp的完整开放阅读框（ORF），其编码721个氨基酸的多肽。在推导的？Sp-HSP90的氨基酸序列中发现了五个定义HSP90蛋白家族的保守区。它在N端包含一个腺苷5'-三磷酸酶（ATPase）结构域，在C-端包含一个保守的签名序列MEEVD。终奌站。实时定量聚合酶链反应（PCR）（qRT-PCR）分析显示？Sp-HSP90？mRNA在不同组织中的分布及其在副溶血弧菌攻击的蟹血细胞中的瞬时表达。在心脏，肝胰腺，肌肉，血细胞，睾丸和卵巢中，除眼球外，均检测到不同水平的？Sp-HSP90？mRNA。活和热灭活细菌攻击后，血细胞中？Sp-Hsp90？mRNA的表达水平明显上调，活细菌组明显高于热灭活细菌组。这些结果表明Sp-HSP90α基因可能作用于S中的免疫力和抗感染性。 paramamosain。

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