《植物生理学报》 2009, 45(12): 1186-1190

山葡萄中类黄酮3’- 羟化酶基因(F3’H) cDNA 的克隆和分析

刘海峰^1,2, 杨成君¹, 赵权¹, 李波¹, 王军^1,*

1 东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨150040; 2 延边大学农学院, 吉林龙井133400

通信作者：王军；E-mail: junwang1966@yahoo.com.cn；Tel: 0451-82191829

摘　要：

用 RT-PCR 和 SMART RACE 技术克隆了山葡萄 F3’H 基因的全长 cDNA 序列。其 GenBank 登陆号为 FJ645766, F3’HcDNA 全长 1 844 bp, 包括开放阅读框1 530 bp, 编码 509 个氨基酸, 该基因表达产物相对分子质量为56.14 kDa, 等电点为8.24, 是不稳定蛋白。该基因属于 P450 超基因家族, 不包含信号肽。氨基酸序列与欧亚种葡萄(AJ8803537)、矢车菊(FJ753550)、金鱼草(DQ272592)、大豆(AB061212)和玉米(EU966228)等植物的F3’H氨基酸序列的同源性系数分别为98%、77%、72%、74% 和 57%。

关键词：山葡萄; cDNA 克隆; 类黄酮 3’- 羟化酶

收稿：2009-09-23   修定：2009-11-09

资助：黑龙江省留学归国人员科学基金(LC08C07)。

cDNA Cloning and Analysis of Flavonoid 3’-hydroxylase (F3’H) in Vitis amurensis Rupr.

LIU Hai-Feng^1,2, YANG Cheng-Jun¹, ZHAO Quan¹, LI Bo¹, WANG Jun^1,*

¹Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, Harbin 150040, China; ²Agricultural College, Yanbian University, Longjing, Jilin 133400, China

Corresponding author: WANG Jun; E-mail: junwang1966@yahoo.com.cn; Tel: 0451-82191829

Abstract:

The full-length cDNA sequences of F3’H genes in Vitis amurensis were cloned by the technique of RT-PCR and SMART RACE. The landing Genbank No. is FJ645766 and the full-length of F3’H cDNA is 1 844 bp with open reading frame of 1 530 bp, which encodes 509 amino acids. The deduced amino acid of F3’H gene was unstable protein with the molecular mass of 56.14 kDa and isoelectric point of 8.24. The F3’H gene belongs to the P450 supergene family, without a signal peptide. The amino acid sequence of F3’H in Vitis amurensis had the homology with F3’H of Vitis vinifera (AJ8803537), Centaurea cyanus (FJ753550), Antirrhinum majus (DQ272592), Glycine max (AB061212) and Zea mays (EU966228), and the homology coefficient was 98%, 77%, 72%, 74% and 57% respectively.

Key words: Vitis amurensis; cDNA cloning; flavonoid 3’-hydroxylase