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刚毛柽柳RING型E3泛素连接酶基因的克隆及表达分析
发表时间：2017-11-15 浏览量：1834 下载量：420
| 全部作者： | 张玉，张悦，张春蕊，王超 |
| 作者单位： | 东北林业大学林木遗传育种国家重点实验室 |
| 摘 要： | E3泛素连接酶是一个种类复杂的蛋白质家族，在对植物生长发育和逆境胁迫的响应等方面起重要的调节作用。通过对刚毛柽柳的转录组数据进行分析，鉴定获得一个E3泛素连接酶基因（ThE3）。ThE3基因cDNA全长2 499 bp，包含1 275 bp开放阅读框（open reading frame，ORF），编码424个氨基酸，编码蛋白分子量45.8 ku，理论等电点（isoelectric point，pI）为8.30. 疏水性分析预测RING型E3泛素连接酶具有较强的亲水性，无信号肽及跨膜区，亚细胞定位于细胞核上。氨基酸多序列比对结果显示，ThE3与荷花的RING-finger类型E3泛素连接酶蛋白序列相似性最高，为76%，且具有RING-finger结构域，初步确定该基因为刚毛柽柳RING-finger类型E3泛素连接酶基因。实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）分析表明，ThE3表达受盐和干旱胁迫诱导，预示ThE3可能在刚毛柽柳适应高盐和干旱环境中发挥重要作用。 |
| 关 键 词： | 分子生物学；刚毛柽柳；RING型E3泛素连接酶；基因克隆；基因表达 |
| Title： | Cloning and expression analysis of RING E3 ubiquitin ligase gene from Tamarix hispida |
| Author： | ZHANG Yu, ZHANG Yue, ZHANG Chunrui, WANG Chao |
| Organization： | State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, |
| Abstract： | The E3 ubiquitin ligase is a complex protein family that plays an important role not only in plant growth and development but also in response to various stresses. A full length cDNA of a RING type E3 ubiquitin ligase (named ThE3) was isolated from the transcriptome cDNA librarys of Tamarix hispida. ThE3 was 2 499 bp in length, including an open reading frame (ORF) of 1 275 bp which was predicted to encode 424 amino acids. The estimated molecular weight and isoelectric point (pI) of the putative protein were 45.8 ku and 8.30, respectively. Hydrophobic analysis predicted that RING type E3 ubiquitin ligase had strong hydrophilicity, no signal peptide and transmembrane region, and subcellular localization on the nucleus. Amino acid multiple sequence analysis showed that ThE3 was 76% similar in amino acid sequence to homology from Nelumbo nucifera. And the RING-finger domain was used to determine the ThE3 as the RING-finger type E3 ubiquitin ligase gene of T. hispida. Quantitative real-time PCR (qRT-PCR) assay revealed that the mRNA levels of ThE3 was significantly up-regulated under NaCl and PEG treatments in T. hispida, suggesting that ThE3 might play an important role in salt and drought tolerance of T. hispida. |
| Key words： | molecular biology; Tamarix hispida; RING type E3 ubiquitin ligase; gene cloning; gene expression |
| 发表期数： | 2017年11月第21期 |
| 引用格式： | 张玉，张悦，张春蕊，等. 刚毛柽柳RING型E3泛素连接酶基因的克隆及表达分析[J]. 中国科技论文在线精品论文，2017，10（21）：2347-2356. |
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