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首页> 外文期刊>Journal of genetics >cDNA cloning and expression analysis of two distinct Sox8 genes in Paramisgurnus dabryanus (Cypriniformes)
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cDNA cloning and expression analysis of two distinct Sox8 genes in Paramisgurnus dabryanus (Cypriniformes)

机译：br形拟南芥中两个不同Sox8基因的cDNA克隆及表达分析

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The Sox9 gene attracts a lot of attention because of its connection with gonadal development and differentiation. However, Sox8, belonging to the same subgroup SoxE, has rarely been studied. To investigate the function as well as the evolutionary origin of SOXE subgroup, we amplified the genomic DNA of Paramisgurnus dabryanu using a pair of degenerate primers. Using rapid amplification of the cDNA ends (RACE), it was discovered that P. dabryanu has two duplicates: Sox8a and Sox8b. Each has an intron of different length in the conserved HMG-box region. The overall sequence similarity of the deduced amino acid of PdSox8a and PdSox8b was 46.26%, and only two amino acids changed in the HMG-box. This is the first evidence showing that there are two distinct duplications of Sox8 genes in Cypriniformes. Southern blot analysis showed only one hybrid band, with lengths 7.4 or 9.2 kb. Both semi-quantitative RT-PCR and real-time quantitative PCR assay displayed that both PdSox8a and PdSox8b are downregulated during early embryonic development. In adult tissues, the two Sox8 genes expressed ubiquitously, and expression levels are particularly high in the gonads and brain. In gonads, both PdSox8a and PdSox8b are expressed at a higher level in the tesis than in the ovary. PdSox8a and PdSox8b may have functional overlaps and are essential for the neuronal development and differentiation of gonads.
机译：Sox9基因由于与性腺发育和分化有关而引起了广泛的关注。然而，很少研究属于同一亚组SoxE的Sox8。为了研究SOXE亚组的功能以及进化起源，我们使用一对简并引物扩增了Paramisgurnus dabryanu的基因组DNA。使用cDNA末端的快速扩增（RACE），发现达氏疟原虫具有两个重复：Sox8a和Sox8b。每个在保守的HMG盒区域中具有不同长度的内含子。 PdSox8a和PdSox8b推导的氨基酸的总体序列相似性为46.26％，并且HMG-box中只有两个氨基酸发生了变化。这是第一个证据表明，在鲤形目中Sox8基因有两个不同的重复。 Southern印迹分析显示仅一个杂合带，长度为7.4或9.2kb。半定量RT-PCR和实时定量PCR分析均显示，PdSox8a和PdSox8b在早期胚胎发育过程中均被下调。在成人组织中，两个Sox8基因无处不在表达，并且在性腺和大脑中的表达水平特别高。在性腺中，PdSox8a和PdSox8b在胚胎中的表达水平高于卵巢。 PdSox8a和PdSox8b可能具有功能重叠，并且对于性腺的神经元发育和分化至关重要。

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