Identification and functional study of a shrimp Dorsal homologue
Identification and functional study of a shrimp Dorsal homologue
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DOI：
10.1016/j.dci.2009.08.009
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发表时间：
2010-02-01
影响因子：
2.9
通讯作者：
He, Jian-Guo
中科院分区：
文献类型：
--
作者：
Huang, Xian-De;Yin, Zhi-Xin;He, Jian-Guo
Rel/NF-kappa B转录因子在诱导和调节先天免疫应答中发挥核心作用。本文报道了凡纳滨对虾（Litopenaeus vannamei）背部同源物LvDorsal的鉴定和功能分析。LvDorsal cDNA全长2204bp，有一个开放阅读框，编码400个氨基酸。推导出LvDorsal含有一个保守的Rel同源结构域（RHD）、一个IPT （ig -样、丛蛋白和转录因子）结构域和一个细胞核定位信号，提示其属于II类NF-kappa b。RT-PCR分析显示，LvDorsal mrna在所有测试组织中均有表达，包括鳃、表皮、红细胞、肠、胃、眼柄、脑、肝胰脏、肌肉、心脏和幽门盲肠。免疫荧光分析显示，重组LvDorsal蛋白被转移到果蝇S2细胞的细胞核中。电泳迁移率转移实验表明，S2细胞的重组LvDorsal RHD与D. melanogaster kappa B基序特异性结合。此外，双荧光素酶报告基因实验表明LvDorsal可以反激活对虾penaeidin-4和果蝇附合蛋白基因51侧翼区控制的报告基因，表明LvDorsal可以调节对虾penaeidin-4基因的转录。对LvDorsal的研究将有助于我们更好地了解对虾的免疫，并可能有助于获得更有效的对虾疾病预防方法。2009爱思唯尔有限公司版权所有。
Rel/NF-kappa B transcription factors play central roles in induction and regulation of innate immune responses. Here, identification and functional analysis of LvDorsal, a Dorsal homologue from the Pacific white shrimp Litopenaeus vannamei, were described. The full-length cDNA of LvDorsal is 2204 bp with an open reading frame that encodes 400 amino acids. The deduced LvDorsal contains a conserved Rel homology domain (RHD), an IPT (Ig-like, plexins and transcription factors) domain and a nucleus localization signal, suggesting that it belongs to the class II NF-kappa B. RT-PCR analysis showed that LvDorsal mRNAs were expressed in all the tissues tested, including gill, epidermis, hemocytes, intestine, stomach, eyestalk, brain, hepatopancreas, muscle, heart and pyloric caecum. Immunofluorescence assay showed that recombinant LvDorsal was translocated into the nucleus of Drosophila S2 cells. Electrophoretic mobility shift assay illustrated that recombinant LvDorsal RHD from S2 cells bound specifically with D. melanogaster kappa B motifs. Additionally, the dual-luciferase reporter assays indicated that LvDorsal could transactivate the reporter gene controlled by the 51 flanking region of shrimp penaeidin-4 and Drosophila attacin genes, suggesting that LvDorsal can regulate the transcription of shrimp penaeidin-4 gene. Study of LvDorsal will help us to better understand shrimp immunity and may help to obtain more effective methods to prevent shrimp diseases. (C) 2009 Elsevier Ltd. All rights reserved.