海蜇(Rhopilema esculentum)even-skipped基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of even-skipped Gene in Jellyfish(Rhopilema esculentum)

Even-skipped(eve)是一个含有同源框结构域的转录调控因子,在体节形成、神经分化、尾芽发育等过程中发挥着重要作用。为探究eve在钵水母变态发育中的作用,本研究以海蜇转录组测序获得的eve基因类似序列为基础,通过RACE技术获得了海蜇eve基因(ReEve)的c DNA全长,并已提交GenBank数据库,登录号为MN836581。结果显示ReEve基因全长1566 bp,开放阅读框(ORF)为630 bp,所编码蛋白质含209个氨基酸,其分子量(MW)为24.29 kD,理论等电点(pI)为8.90。在二级结构上,无规则卷曲为ReEve蛋白最主要的结构形式。ReEve蛋白是一个亲水性蛋白质,不存在跨膜区域,不含信号肽。在第76~135位氨基酸之间具有eve相关基因家族特有的同源框结构域(homeodomain),与其他物种具有较高同源性。基于氨基酸序列获得的系统进化关系显示与传统分类结果一致。研究结果为进一步揭示其在海蜇等钵水母横裂生殖中的具体作用提供了一定基础。

Even-skipped(eve)is a homeodomain containing transcription factor,which plays an important role in various development processes,such as segmentation,neurogenesis,tailbud formation and so on.To explore the role of eve in the metamorphosis of scyphozoan jellyfish,full length cDNA sequence of eve gene from Rhopilema esculentum(ReEve)was obtained by rapid amplification of cDNA ends(RACE)based on the RNA-seq derived eve-like sequence fragment.It has been submitted to the GenBank database and the accession number is MN836581.The results showed that the full length c DNA of ReEve was 1566 bp,includingan open reading frame(ORF)of 630 bp length and encoding 209 amino acids.The molecular weight(MW)and theoretical isoelectric point(pI)were predicted to be 24.29 kD and 8.90 respectively.In the secondary structure,the random coil was the main structural form of the protein.ReEve was a hydrophilic protein containing none transmembrane structure and signal peptides.R eEve had a specific homeodomain of eve related gene family between the 76th to135th amino acid,which showed great identity to the eve gene of other species.The phylogenetic relationship revealed by the amino acid sequences was consistent with the traditional taxonomy.The results offered a foundation for uncovering the exact role of eve gene in the strobilation of Rh.esculentum and other scyphozoan.