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利用RACE的方法从麝香百合‘白天堂’叶片中克隆出热激转录因子(Heat shock transcription factor,Hsf)A1家族中一个成员的cDNA全长序列。该基因编码序列长为1 587 bp,编码528个氨基酸,蛋白质分子量为59.056 kD。序列和进化树分析表明,该基因具有A1类Hsf的5个关键功能域和调控基序,与水稻的OsHsfA1a氨基酸序列同源性最高,说明该基因为HsfA1家族的新成员,将其命名为LlHsfA1。利用荧光定量PCR的方法分析LlHsfA1表达模式,结果表明常温下其可在根、鳞茎和叶片中表达,说明该基因为组成型表达;42℃热激处理1~12 h,该基因的表达量在叶片中呈现“上升—下降”交替变化的趋势。亚细胞定位表明,LlHsfA1定位在细胞核和细胞质中。
The full - length cDNA sequence of one member of heat shock transcription factor (Hsf) A1 family was cloned from leaves of Musk lily ’BaiTian’ by RACE. The coding sequence of this gene was 1 587 bp, encoding a protein of 528 amino acids with a molecular mass of 59.056 kD. Sequence and phylogenetic tree analysis showed that the gene has five key functional domains and regulatory motifs of class A1 Hsf, which has the highest homology with the OsHsfA1a amino acid sequence of rice, indicating that the gene is a new member of HsfA1 family and named as LlHsfA1 . The expression pattern of LlHsfA1 was analyzed by fluorescence quantitative PCR. The results showed that the gene was expressed constitutively in roots, bulbs and leaves at room temperature, indicating that the gene was constitutively expressed at 42 ℃ for 1 to 12 h, Leaves showed “rising - falling” alternating trend. Subcellular localization showed that LlHsfA1 is localized in the nucleus and cytoplasm.