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园艺学报 ›› 2012, Vol. 39 ›› Issue (11): 2199-2205.

• 观赏植物 • 上一篇    下一篇

百合热激转录因子基因LlHSF1 的克隆与表达分析

 易瑾, 罗弦, 曹兴, 陈莉, 辛海波, 李晓昕, 陈进, 义鸣放   

  1. (中国农业大学观赏园艺与园林系,北京 100193)
  • 出版日期:2012-11-25 发布日期:2012-11-25

Cloning and Expression Analysis of LlHSF1 from Lilium longiforum

 YI  Jin, LUO  Xian, CAO  Xing, CHEN  Li, XIN  Hai-Bo, LI  Xiao-Xin, CHEN  Jin, YI  Ming-Fang   

  1. (Department of Ornamental Horticulture and Landscape Architecture,China Agricultural University,Beijing 100193,China)
  • Online:2012-11-25 Published:2012-11-25

摘要: 以铁炮百合(Lilium longiforum)‘白天堂’的组培苗叶片为试材,通过RACE方法得到了一个热激转录因子(heat shock transcription factor,HSF)基因LlHSF1的cDNA全长序列。该序列全长1 068 bp,推断其含有一个780 bp的开放阅读框(ORF),编码260个氨基酸,推导的蛋白质分子量为30.123kD。对推定的氨基酸序列与其它已知物种HSF进行比对发现,该基因有热激转录因子所具备的典型结构域和调控元件,据此推断,克隆到的基因是一个新的热激转录因子成员,命名为LlHSF1。荧光定量PCR分析结果表明:常温下该基因在百合根、鳞茎、叶中都有表达,但在叶片中表达量较高;实时荧光定量PCR分析结果表明,42℃处理1 ~ 12 h,叶中该基因表达量增加。

关键词: 百合, 铁炮百合, 热激转录因子, 克隆, 表达, 荧光定量RT-PCR

Abstract: A full length cDNA named LlHSF1,belonging to Heat shock transcription factor(HSF),was cloned from Lilium longiforum‘White Heaven’by RT-PCR and RACE(rapid amplification of cDNA ends). The gene consisted of 1 068 bp and open reading frame encompassed 780 bp encoding a polypeptide of 260 amino acids with calculated protein molecular mass of 30.123 kD. Cluster analysis showed that this gene was highly homologous to known HSF from other organisms. RT-PCR analysis showed that expression of LlHSF1 could be detected in leaf,root and bulb at 22 ℃,and it was expressed highly in the leaf.Theresults with SYBR GREEN real-time quantitative PCR showed that the gene expression level increased in leaves during 1–12 hours at 42 ℃.

Key words: Lilium longiforum, heat shock transcription factor(HSF), cloning, real-time quantitative PCR