百合热激转录因子基因LlHSF1的克隆与表达分析

Cloning and Expression Analysis of LlHSF1 from Lilium longiforum

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1068bp,推断其含有一个780bp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1～12h,叶中该基因表达量增加。

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 ℃.