木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因克隆与序列分析  

Cloning and bioinformatics analysis of VvGST1 from Munage tab.grapes

作　　者：王曼 张政 伊丽达娜·迪力夏提 吴斌 WANG Man;ZHANG Zheng;Yilidana Dilixiati;WU Bin(College of Food and Pharmaceutical Sciences,Xinjiang Agricultural University,Urumqi 830052,China;Institute of Agro-products Storage and Processing,Xinjiang Academy of Agricultural Sciences/Xinjiang Key Laboratory of Processing and Preservation of Agro-products,Urumqi 830091,China)

机构地区：[1]新疆农业大学食品科学与药学学院,乌鲁木齐830052 [2]新疆农业科学院农产品贮藏加工研究所/新疆农产品加工与保鲜重点实验室,乌鲁木齐830091

出　　处：《新疆农业科学》2023年第8期1922-1930,共9页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(U2003213);国家科学自然基金项目(31860460);新疆维吾尔自治区区域协同创新专项(2021E01005)。

摘　　要：【目的】克隆木纳格葡萄果实中谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)基因VvGST1全长并研究其序列特征,为该基因在葡萄果实抗病作用和功能的研究奠定基础。【方法】根据已有的基因序列,设计3'和5'端RACE引物,利用RT-PCR和RACE技术克隆VvGST1基因cDNA全长。【结果】该基因序列全长719 bp,均为开放阅读框(ORF),共编码221个氨基酸。该基因编码蛋白相对分子质量为25.55 kDa;理论等电点pI值为6.32;分子式为C_(1178)H_(1799)N_(293)O_(321)S_(11);不稳定指数为39.22;该蛋白质是稳定的亲水性蛋白,不含跨膜结构域,没有预测到信号肽,可能存在于细胞基质中,是典型的基质蛋白。VvGST1氨基酸序列与棉花、桃、西梅、樱桃、李子、板栗等植物聚为一类,其中与棉花属亲缘关系最近。【结论】获得了木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因全长编码序列,探明了该序列的结构特征。【Objective】To clone and study the sequence characteristics of the glutathione-S-transferase,(GST)gene from the Vitis vinifera L.cv‘Munage’table grapes in the hope of laying a foundation for the disease resistance and function of this gene in grape fruit.【Methods】Full-length VvGST1 gene cDNA was isolated from grape fruit according to the existed ESTs sequence,and 3'and 5'end of the RACE primers were designed by using RT-PCR and RACE.【Results】A variety of bioinformatics analysis methods were used to predict and analyze the gene,and the results showed that the ORF(Open Reading Frame)length was 719 bp,which encoded a 221 amino acid polypeptide.The VvGST1 gene was forecasted and the relative molecular mass of encoding the protein was 25.55 kDa,the theoretical isoelectric point was 6.32,the atomic composition was C_(1178)H_(1799)N_(293)O_(321)S_(11).Its instability coefficient was 39.22,showing it was astable hydrophilic protein without signal peptide or transmembrane structure.It might exist in the cell matrix and was a typical matrix protein.Through analyzing the evolutionary tree,the amino acid sequence of VvGST1 was clustered with cotton,peach,prunus,cherry,plum,chestnut and other plants,among which it was closely related to cotton.【Conclusion】The full-length coding region of VvGST1 gene in Vitis vinifera L.cv‘Munage’table grapes is successfully obtained,and the structural characteristics of the sequence are confirmed.

关 键 词：葡萄 克隆 谷胱甘肽-S-转移酶 VvGST1基因 序列分析 

分 类 号：S663.1[农业科学—果树学]