Molecular cloning and expression of a TNF receptor and two TNF ligands in the fish ovary
Molecular cloning and expression of a TNF receptor and two TNF ligands in the fish ovary
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DOI：
10.1016/s1096-4959(01)00353-0
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发表时间：
2001-06-01
影响因子：
2.2
通讯作者：
Goetz, FW
中科院分区：
文献类型：
--
作者：
Bobe, J;Goetz, FW
使用变性引物，通过PCR斑马鱼和鳟鱼cDNA或cDNA文库获得TNF（肿瘤坏死因子）受体和两个TNF配体的部分cDNA。然后用这些片段筛选合适组织的cDNA文库，以获得含有完整编码序列的克隆。获得了一个斑马鱼cDNA，推测其编码438个氨基酸的卵巢TNF受体（OTR），该受体被鉴定为TNF受体家族的含死亡结构域的成员。在北方印迹中，OTR cDNA与3.4-kb转录物杂交，该转录物在斑马鱼卵巢中丰富，但在所有其他测试组织中轻微检测到。一个斑马鱼的cDNA可能编码214个氨基酸的蛋白质与哺乳动物TRAIL（肿瘤坏死因子相关的凋亡诱导配体）的序列相似性，也被分离出来。此外，获得了溪鳟TRAIL同源物的片段。最后，全长溪鳟cDNA，推测编码255个氨基酸的蛋白质与哺乳动物TNF-α和α-光毒素的序列相似性，被分离。这项研究是第一次报告的死亡结构域含有TNF受体和第一次发表的报告的TNF配体在鱼类。(C)2001 Elsevier Science Inc. All rights reserved.
Using degenerative primers, partial cDNAs of a TNF (tumor necrosis factor) receptor and two TNF ligands were obtained by PCR of zebrafish and trout cDNAs, or cDNA libraries. These fragments were then used to screen cDNA libraries of appropriate tissues to obtain clones containing full coding sequences. A zebrafish cDNA was obtained that presumably codes for a 438 amino acid ovarian TNF receptor (OTR) that was identified as a death-domain-containing member of the TNF receptor family. On Northern blots, the OTR cDNA hybridized with a 3.4-kb transcript that is abundant in the zebrafish ovary but lightly detected in all other tissues tested. A zebrafish cDNA presumably coding for a 214 amino acid protein with sequence similarity to mammalian TRAIL (TNF-related apoptosis inducing ligand), was also isolated. In addition, a fragment of the brook trout TRAIL homologue was obtained. Finally, a full-length brook trout cDNA, that presumably codes for a 255 amino acid protein with sequence similarity to mammalian TNF-alpha and lymphotoxin-alpha, was isolated. This study is the first report of a death-domain-containing TNF receptor and the first published report of a TNF ligand in fish. (C) 2001 Elsevier Science Inc. All rights reserved.