向日葵E3泛素连接酶基因克隆及表达分析

Cloning and Expression Analysis of E3 Ubiquitin Ligase Gene HERC2 of Sunflower(Helianthus annuus L.)

该研究从向日葵中克隆了E3泛素连接酶基因HERC2,并进行了生物信息学分析和不同胁迫条件的表达分析。序列分析表明,HERC2(登录号为KT832066)序列的CDS为1 608bp,编码535个氨基酸,预测其分子量131kD,等电点为5.03。HERC2编码的蛋白质为疏水性蛋白质,且为细胞质蛋白;亚细胞定位预测分析表明,向日葵HERC2可能定位在高尔基体中;该蛋白质有5个RCC1保守结构域。向日葵HERC2与已报道的其他植物同源蛋白有相似的保守区域,与醉蝶花亲缘关系最近,而与大豆和野生大豆的亲缘关系最远。与HERC2cDNA对应的gDNA(登录号为KT832067)的ORF长度为3 409bp,与cDNA编码序列比对结果表明,该gDNA由5个外显子和4个内含子组成。实时荧光定量PCR分析表明,向日葵HERC2基因表达受非生物胁迫调节,在不同器官及不同非生物胁迫下存在特异性表达差异。研究认为,HERC2基因应答逆境胁迫具有其特定的表达模式,研究结果为加强对HERC2的利用奠定了基础。

In order to understand the functions of HERC2 and effectively make use it, we cloned the HERC2 cDNA and gDNA sequence from sunflower and analyzed it by bioinformatics method. Expression pattern of HERC2 of sunflower was revealed under abiotic stresses. The CDS of HERC2 is 1 608 bp, encoding 535 amino acids. The predicted molecular weight is 131 kD, isoelectric point is 5.03. The HERC2 encoded protein is hydrophobic proteins, and it is cytoplasmic protein. The protein was localized in the Golgi apparatus. The protein has 5 RCC1 conserved domains. Phylogenetic analysis of amino acid sequence of E3 ubiquitin ligase was close to Tarenaya hassleriana, was far with Glycine max and Glycine soja. Gene accession number is KT832066. The ORF of gDNA of HERC2 is 3 409 bp, which have 5 exons and 4 introns. Gene accession number is KT832067. RT-qPCR showed that HERC2 has different expression characteristics under different abiotic stresses. The expression characteristics also exist in different organs.