Molecular cloning and responsive expression of macrophage expressed gene from small abalone Haliotis diversicolor supertexta
Molecular cloning and responsive expression of macrophage expressed gene from small abalone Haliotis diversicolor supertexta
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DOI：
10.1016/j.fsi.2007.12.008
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发表时间：
2008-03-01
影响因子：
4.7
通讯作者：
Wang, Yi-Lei
中科院分区：
文献类型：
--
作者：
Wang, Guo-Dong;Zhang, Ke-Feng;Wang, Yi-Lei
采用同源克隆和cDNA末端快速扩增（RACE）技术，从九孔鲍（Haliotis diversicolor supertexta）中克隆了穿孔素样分子巨噬细胞表达基因（saMpeg 1）的cDNA全序列。saMpeg 1的cDNA全长为2781 bp，由252 bp的5 '端非翻译区（UTR）、342 bp的3'端非翻译区（UTR）和2184 bp的开放阅读框组成，其中3 '端UTR含有信号序列TAA和poly（A）尾。推导的蛋白质（saMpeg 1）由728个氨基酸组成，并含有穿孔素的溶细胞“螺旋-转角-螺旋”结构域（残基171-218），其中的α-螺旋是两亲性的穿孔素。一个推定的单跨膜结构域位于残基667-689处，紧接着是一个修饰的弗林蛋白酶切割位点（KRRRK;残基689-693）。真实的时间定量PCR结果显示，在接种副溶血性弧菌后8 h和96 h，saMpeg 1基因有较高的表达，而TBT暴露后，saMpeg 1基因表达无明显变化。与哺乳动物穿孔素的结构相似性以及对细菌感染和TBT暴露的不同基因表达水平表明saMpeg 1可能在小鲍抗微生物的免疫应答中发挥作用。(C)2008爱思唯尔有限公司保留所有权利。
The complete cDNA sequence of macrophage expressed gene (saMpeg1), a perforin-like molecule, was isolated from small abalone (Haliotis diversicolor supertexta) by homology cloning and rapid amplification of cDNA ends (RACE). The full-length cDNA of saMpeg1 was 2781 bp, consisting of a 5'-terminal untranslated region (UTR) of 252 bp, a 3'-terminal UTR of 342 bp with a signal sequence TAA and a poly (A) tail, and an open reading frame of 2184 bp. The deduced protein (saMpeg1) was composed of 728 amino acids, and contains the cytolytic "helix-turn-helix" domain of perforin (residues 171-218), of which the alpha-helices are amphipathic as are those of perforin. A putative single transmembrane domain is located at residues 667-689, and a modified furin cleavage site (KRRRK; residues 689-693) immediately follows. The result of real time quantitative PCR showed that saMpeg1 was highly expressed at 8 h and 96 h post-injection of the Gram-negative bacterium Vibrio parahaemolyticus, but there was no change after TBT exposure. The structural similarity to mammalian perforin and the different gene expression level to bacterial infection and TBT exposure suggest that saMpeg1 may play a role in the immune response against microorganisms in small abalone. (C) 2008 Elsevier Ltd. All rights reserved.