东方山羊豆蔗糖转运蛋白基因克隆及表达载体构建

摘要

Based on an EST sequence of Galega orientalis, a full length of 1883bp cDNA was isolated by using RACE technology, termed GoSUT (GenBank accession No. HM802255). The sequence analysis revealed that GoSUT contains an open reading frame (ORF) of 1542bp and encodes 514 amino acids residues. The protein function and structure were analyzed using Bioinformatics tools. Phylogenetic tree analysis indicated that GoSUT belongs to the type Ⅰ SUT. RT-qPCR indicated that the expression of GoSUT gene was stronger in stem than in root and leaf. Under PEG and NaCl stress, relative gene expression quantity was increased a little. The plant expression vector 1302-GoSUT was constructed by the aid of pCAMBIA-1302 vector.%根据已知的EST序列,采用RACE技术克隆了东方山羊豆蔗糖转运蛋白基因全长eDNA,命名为GoSUT(GenBank accession No.HM802255).序列分析结果表明,该cDNA全长1883bp,开放阅读框1545bp,编码514个氨基酸.应用生物信息学软件对编码蛋白的理化性质、结构和功能进行了分析和预测.进化树分析显示,该蛋白属于蔗糖转运蛋白第Ⅰ类.利用Real-Time PCR方法分析表明,GoSUT在茎中表达量高,在根、叶中表达量低.在PEG、NaCl胁迫处理下,GoSUT基因相对表达量都有不同程度上调.利用pCAMBIA1302质粒,成功构建了GoSUT基因植物表达载体.