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日本沼虾钙调素依赖蛋白激酶 I 的 cDNA 克隆及在蜕皮中的作用

Calcium-calmodulin dependent protein kinase I from Macrobrachium nipponense: cDNA cloning and involvement in molting.

机构信息

Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, PR China; Wuxi Fisheries College, Nanjing Agricultural University, Nanjing 210095, PR China.

Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, PR China; Wuxi Fisheries College, Nanjing Agricultural University, Nanjing 210095, PR China.

出版信息

Gene. 2014 Apr 1;538(2):235-43. doi: 10.1016/j.gene.2014.01.055. Epub 2014 Jan 31.

Abstract

Calcium-calmodulin dependent protein kinase I is a component of a calmodulin-dependent protein kinase cascade and involved in many physiological processes. The full-length cDNA of calcium-calmodulin dependent protein kinase I (MnCaMKI) was cloned from the freshwater prawn Macrobrachium nipponense and its expression pattern during the molt cycle and after eyestalk ablation is described. The full-length cDNA of MnCaMKI is 3,262 bp in length and has an open reading frame (ORF) of 1,038 bp, encoding a 345 amino acid protein. The expression of MnCaMKI in three examined tissues was upregulated in the premolt stage of the molt cycle. Its expression was induced after eyestalk ablation (ESA): the highest expression level was reached 1 day after ESA in hepatopancreas, and 3 days after ESA in muscle. By dsRNA-mediated RNA interference assay, expression of MnCaMKI and ecydone receptor gene (MnEcR) was significantly decreased in prawns treated by injection of dsMnCaMKI, while expression of these two genes was also significantly decreased in prawns treated by injection of dsMnEcR, demonstrating a close correlation between the expression of these two genes. These results suggest that CaMKI in M. nipponense is involved in molting.

摘要

钙调素依赖性蛋白激酶 I 是钙调素依赖性蛋白激酶级联反应的一个组成部分,参与许多生理过程。我们从日本沼虾(Macrobrachium nipponense)中克隆了钙调素依赖性蛋白激酶 I(MnCaMKI)的全长 cDNA,并描述了其在蜕皮周期和眼柄切除后的表达模式。MnCaMKI 的全长 cDNA 长 3262 个碱基对,具有一个 1038 个碱基对的开放阅读框(ORF),编码一个 345 个氨基酸的蛋白质。在三个检查的组织中,MnCaMKI 的表达在蜕皮周期的前蜕皮期上调。眼柄切除(ESA)后其表达被诱导:在肝胰腺中,ESA 后 1 天达到最高表达水平,在肌肉中,ESA 后 3 天达到最高表达水平。通过 dsRNA 介导的 RNA 干扰试验,注射 dsMnCaMKI 的对虾中 MnCaMKI 和蜕皮激素受体基因(MnEcR)的表达显著降低,而注射 dsMnEcR 的对虾中这两个基因的表达也显著降低,表明这两个基因的表达密切相关。这些结果表明,日本沼虾中的 CaMKI 参与蜕皮。

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