山葡萄谷胱甘肽S-转移酶基因(VAmGST4)克隆及表达分析

刘海峰1,2, 王军1,*
1中国农业大学食品科学与营养工程学院葡萄与葡萄酒研究中心, 北京100083; 2延边大学农学院, 吉林延吉133000

通信作者:王军;E-mail: jun_wang@cau.edu.cn;Tel: 010-62738658

摘 要:

采用RT-PCR和SMART RACE技术克隆了山葡萄VAmGST4基因的全长cDNA序列, GenBank登录号为FJ645770, 基因 全长885 bp, 包括开放阅读框(ORF) 642 bp, 编码213个氨基酸。该基因表达产物相对分子质量为24.24 kDa, 等电点为5.72, 是不稳定蛋白; 该基因属于GST超基因家族, 不包含信号肽。氨基酸序列与欧亚种葡萄(AY971515)、荔枝(EF613493)、矮 牵牛(Y07721)、紫苏(AB362191)和玉米(EU964162)等植物的GST氨基酸序列的同源性系数分别为99%、67%、64%、61% 和47%。半定量PCR显示, 在果实着色过程中, VAmGST4在果皮中表达随花色苷的合成上调; 在茎、果肉和果皮中均有表 达, 而在叶片中不表达, 表明VAmGST4的表达与花色苷的生物合成密切相关。

关键词:山葡萄; cDNA克隆; 谷胱甘肽S-转移酶

收稿:2011-08-01   修定:2011-10-25

资助:现代农业产业技术体系专项资金(CARS-30)。

cDNA Cloning and Expression Analysis of Glutathione S-Transferase (VAmGST4) in Vitis amurensis Rupr.

LIU Hai-Feng1,2, WANG Jun1,*
1Center for Viticulture and Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; 2Agricultural College of Yanbian University, Yanji, Jilin 133000, China

Corresponding author: WANG Jun; E-mail: jun_wang@cau.edu.cn; Tel: 010-62738658

Abstract:

The full-length cDNA sequences of VAmGST4 gene in Vitis amurensis were cloned by the technique of RT-PCR and SMART RACE. The landing Genbank No. is FJ645770 and the full-length of VAmGST4 cDNA is 885 bp with open reading frame of 642 bp, which encodes 213 amino acids. The product of VAmGST4 gene expression is unstable protein with the molecular mass of 24.24 kDa, isoelectric point of 5.72. The VAmGST4 gene belongs to the GST supergene family, not containing a signal peptide. The amino sequence of GST in Vitis amurensis has the homology with GST of Vitis vinifera (AY971515), Litchi chinensis (EF613493), Petunia hybrida (Y07721), Perilla frutescens (AB362191) and Zea mays (EU964162), and the homology coefficient is 99%, 67%, 64%, 61% and 47% respectively. Semi-quantitative PCR analysis showed that VAmGST4gene was expressed in the stem, flesh and skin except in the leaf. Particularly, during coloring process, the expression level of VAmGST4 in skin increased progressively by the accumulation of anthocyanins. The result demonstrated that the expression level of VAmGST4 had close correlation with the accumulation of anthocyanins.

Key words: Vitis amurensis; cDNA cloning; glutathione S-transferase

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