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中华蜜蜂硫氧还蛋白还原酶 1 基因的克隆、结构特征及表达分析。

Cloning, structural features, and expression analysis of the gene encoding thioredoxin reductase 1 from Apis cerana cerana.

机构信息

College of Life Sciences, Shandong Agricultural University, Taian, Shandong, 271018, PR China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2010 Jul;156(3):229-36. doi: 10.1016/j.cbpb.2010.04.003. Epub 2010 Apr 14.

Abstract

Previous studies have demonstrated that mammalian thioredoxin reductases 1 (TrxRs; EC 1.6.4.5) play roles in protection against oxidant injury, cell growth and transformation, and the recycling of ascorbate from its oxidized form. However, little is known about the function of TrxRs in insects, especially in Apis cerana cerana. To gain a better understanding of its role in insects, we cloned TrxR1 from A.cerana cerana (AccTrxR1) and investigated its structural features and expression. The full-length cDNA is 1998 bp long and contains an open reading frame (ORF) of 1485 bp. The deduced AccTrxR1 protein has 494 aa, a calculated molecular mass (MW) of 54.24 kDa, and an isoelectric point of pH 7.35. The full-length genomic DNA of AccTrxR1 is 3500 bp and contains eight exons and seven introns. In addition, the sequence and putative transcription factor-binding sites of the 5'-flanking region were examined. We also looked for conserved domains/motifs, generated homologous alignments, conducted phylogenetic analysis, and made secondary and tertiary structure predictions using web software. RT-PCR revealed that the expression of AccTrxR1 could be induced by UV and heat (37 degrees C). These results indicate that AccTrxR1 may play a key role in protection against oxidant stress.

摘要

先前的研究表明哺乳动物硫氧还蛋白还原酶 1(TrxRs;EC 1.6.4.5)在对抗氧化剂损伤、细胞生长和转化以及抗坏血酸从其氧化形式的再循环方面发挥作用。然而,关于 TrxRs 在昆虫中的功能知之甚少,特别是在中华蜜蜂(Apis cerana cerana)中。为了更好地了解其在昆虫中的作用,我们从中华蜜蜂(A.cerana cerana)中克隆了 TrxR1(AccTrxR1),并研究了其结构特征和表达。全长 cDNA 长 1998 bp,包含一个 1485 bp 的开放阅读框(ORF)。推断的 AccTrxR1 蛋白含有 494 个氨基酸,计算的分子量(MW)为 54.24 kDa,等电点为 pH 7.35。AccTrxR1 的全长基因组 DNA 为 3500 bp,包含 8 个外显子和 7 个内含子。此外,还检查了 5'-侧翼区域的序列和假定转录因子结合位点。我们还寻找了保守结构域/基序,进行了同源比对,进行了系统发育分析,并使用网络软件进行了二级和三级结构预测。RT-PCR 显示 AccTrxR1 的表达可以被 UV 和热(37 度)诱导。这些结果表明 AccTrxR1 可能在对抗氧化剂应激中发挥关键作用。

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