Molecular cloning, expression, and functional analysis of caspase-10 from Japanese flounder Paralichthys olivaceus.
Molecular cloning, expression, and functional analysis of caspase-10 from Japanese flounder Paralichthys olivaceus.
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DOI：
10.1016/j.fsi.2007.07.001
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发表时间：
2007-12
影响因子：
4.7
通讯作者：
T. Kurobe;I. Hirono;H. Kondo;M. Yamashita;T. Aoki
中科院分区：
文献类型：
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作者：
T. Kurobe;I. Hirono;H. Kondo;M. Yamashita;T. Aoki
本研究分离并测序了牙鲆caspase-10基因的全长序列。该基因全长2282bp，编码495个氨基酸残基。在JF-caspase-10以及三个天冬氨酸残基(D-186、-382和-392)中观察到caspase的特征死亡效应结构域(DED)，它们是大亚基和小亚基结构的潜在切割位点。参与催化活性的氨基酸残基(His-325)和五肽(QACQG)在牙鲆caspase-10中是绝对保守的。JF-caspase-10基因全长6.6kb，由11个外显子和10个内含子组成，与人类相似。JF-caspase-10mRNA在鳃、外周血白细胞、脾和后肾中表达较强，在头肾、心、肠、皮肤和胃中表达较弱。TUNEL法检测JF-caspase-10基因在牙鲆细胞系HINAE中的过表达，结果表明，JF-caspase-10基因在转染后24小时可诱导细胞发生凋亡。
We isolated and sequenced caspase-10 cDNA and gene from Japanese flounder, Paralichthys olivaceus. The Japanese flounder (JF)-caspase-10 cDNA consisted of 2282bp and encoded 495 amino acid residues. The characteristic death effector domains (DEDs) of caspases were observed in JF-caspase-10 as well as the three aspartic acid residues (D-186, -382 and -392), which are potential cleavage sites for the large and small subunit structures. The amino acid residue (His-325) and pentapeptide (QACQG), which are involved in catalytic activity, were absolutely conserved in Japanese flounder-caspase-10. JF-caspase-10 gene has a length of 6.6kb and consists of 11 exons and 10 introns similar to that of human. The strong expression of JF-caspase-10 mRNA was detected in the gills, peripheral blood leukocytes, spleen and posterior kidney, while the weak expression was observed in the head kidney, heart, intestine, skin and stomach. The over-expression analysis of JF-caspase-10 in Japanese flounder cell line HINAE was shown to induce apoptosis 24h post-transfection using TUNEL assay.