肿瘤坏死因素联系受体的因素(TRAF2 ) 2 是一个关键部件几乎发信号的全部肿瘤坏死因素受体总科小径。在现在的学习, TRAF2 基因从草鲤鱼被克隆 { Ctenopharyngodon idella ) 由反向的抄写聚合酶,链反应和 cDNA 的快速的扩大结束。全身的 cDNA 是 3162 bp ,包括 60 bp 5' untranslated 区域( UTR ), 1611 bp 开的读框架,和 1491 bp 3' UTR.The polyadenylation 信号( AATAAA )和 mRNA 不稳定性主题( ATTTTA , ATTTA )被跟随由一 poly ,在3' UTR.No 信号肽或 transmembrane 区域的(A)尾巴在草鲤鱼 TRAF2 ( gcTRAF2 )的通常认为的氨基酸被发现了。种系发生的树分析清楚地证明 gcTRAF2 几乎金鱼的 TRAF2 基因。有它在从 56% ~ 97%.It 的另外的脊椎动物范围的相当或相同的事物的 gcTRAF2 的身份被一戒指类型签名在 N 终点,在中间的部分的一根锌手指,和由组成的一个保存 TRAF 领域描绘一 C 近似(TRAF-C ) 子域并且一 N 近似(TRAF-N ) 子域。在在脊椎动物的所有 TRAF2 相当或相同的事物之中的 TRAF-C 的身份从 78% ~ 97% 变化,而从 56% ~ 100%.The recombinant gcTRAF2 的 TRAF-N 范围的身份用 pET-32a 表示向量在 Escherichia 关口 i 被表示了。兔子 anti-gcTRAF2 polyclonal 抗体被获得。在不同机关的 gcTRAF2 的表示被即时量的聚合酶链反应和蛋白质印迹分析检验。它广泛地在心,头肾,胸腺,大脑,鳃,肝,怒气,和箱子肾被散布。这是在鱼的一个 TRAF2 相当或相同的事物分子的第一份报告。
Tumor necrosis factor receptor-associated factor 2 (TRAF2) is a crucial component of almost the entire tumor necrosis factor receptor superfamily signaling pathway. In the present study, a TRAF2 gene has been cloned from grass carp (Ctenopharyngodon idella) by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The full-length cDNA is 3162 bp, including a 60 bp 5' untranslated region (UTR), a 1611 bp open reading frame, and a 1491 bp 3' UTR. The polyadenylation signal (AATAAA) and the mRNA instability motifs (ATTTTA, ATTTA) were followed by a poly(A) tail in the 3' UTR. No signal peptide or transmembrane region has been found in the putative amino acids of grass carp TRAF2 (gcTRAF2). Phylogenetic tree analysis clearly showed that gcTRAF2 is nearest to the TRAF2 gene of goldfish. The identity of gcTRAF2 with its homologs in other vertebrates ranges from 56% to 97%. It is characterized by one RING-type signature at the N-terminus, one zinc finger in the middle part, and one conserved TRAF domain consisting of a C-proximal (TRAF-C) subdomain and a N-proximal (TRAF-N) subdomain. The identity of TRAF-C among all TRAF2 homologs in vertebrates varies from 78% to 97%, whereas the identity of TRAF-N ranges from 56% to 100%. The recombinant gcTRAF2 has been expressed in Escherichia coli using pET-32a expression vector. The rabbit anti-gcTRAF2 polyclonal antibody was obtained. The expression of gcTRAF2 in different organs was examined by real-time quantitative polymerase chain reaction and Western blot analysis. It was widely distributed in heart, head kidney, thymus, brain, gill, liver, spleen, and trunk kidney. This is the first report of a TRAF2 homolog molecule in fish.