能源植物橡胶草TkGGPPS基因的克隆及表达分析

李永梅, 冯玉杰, 李锦, 赵李婧, 闫洁*
石河子大学生命科学学院, 新疆石河子832000

通信作者:闫;E-mail: jiey@shzu.edu.cn

摘 要:

牻牛儿基牻牛儿基焦磷酸合酶(geranylgeranyl pyrophosphate synthase, GGPPS)是植物细胞萜类物质合成的重要调节靶点。利用RT-PCR和RACE技术首次从橡胶草(Taraxacum kok-saghyz)中克隆得到一个GGPPS基因, 命名为TkGGPPS (Gen-Bank: KT028713), TkGGPPS cDNA全长1 140 bp, 编码379个氨基酸。序列分析表明, TkGGPPS基因属于类异戊二烯超家族成员, 其功能可能与萜类物质的合成有关。氨基酸序列同源性比对与系统进化分析表明, TkGGPPS与向日葵HaGGPPS的亲缘关系较近, 相似性为74.67%。实时定量PCR的结果表明, TkGGPPS基因在第6周橡胶草的主根和成熟的种子中显著表达。上述结果为进一步研究橡胶草产胶代谢途径和产胶合成相关基因功能提供一定的理论指导和技术支持。

关键词:橡胶草; GGPPS; 基因克隆; 序列分析; 表达分析

收稿:2015-11-18   修定:2016-01-11

资助:国家自然科学基金(31360060)。

Cloning and expression analysis of the TkGGPPS gene from engery plant Taraxacum kok-saghyz

LI Yong-Mei, FENG Yu-Jie, LI Jin, ZHAO Li-Jing, YAN Jie*
College of Life Sciences, Shihezi University, Shihezi, Xinjiang 832000, China

Corresponding author: YAN Jie; E-mail: jiey@shzu.edu.cn

Abstract:

Geranylgeranyl pyrophosphate synthase (GGPPS) is one of the important regulating targets in plant cell terpenoid substances synthetic. A new cDNA sequence encoding GGPPS, designated as TkGGPPS (Gen-Bank: KT028713), from Taraxacum kok-saghyz was cloned by RT-PCR and RACE techniques and assayed by bioinformatics. The cDNA sequence of TkGGPPS was of 1 140 bp encoding 379 amino acids. Sequence analysis showed that TkGGPPS was one of the members of short chain prenyltransferases super family. Its function may be associated with the synthesis of terpenoid substances. Amino acid sequence homology alignment and phylogenetic analysis showed that the similarition of TkGGPPS and HaGGPPS from Helianthus annuus were more than 74.67%. Quantitative RT-PCR analysis revealed that TkGGPPS genes expressed in 6th-week T. koksaghyz root and mature seeds significantly. This work will provide certain theoretical guidance and technical support for further study to metabolic pathways and functional gene related to the synthetic rubber.

Key words: Taraxacum kok-saghyz; GGPPS; gene clone; sequence analysis; expression analysis

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