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以香樟(Cinnamomum camphora)实生苗为试验材料,利用同源克隆结合RACE技术获得了两个冷诱导转录因子基因CBF/DREB1的cDNA全长序列,命名为CcCBFc和CcCBFd,Gen Bank登录号分别为KP336741和KP336742。序列分析显示这两个基因均没有内含子,cDNA全长为897和1 010 bp,开放阅读框分别为654和621 bp,编码217和206个氨基酸,预测蛋白分子量分别为24.0和22.9 kD,等电点分别为5.26和8.58。基于氨基酸序列的同源性比对和系统进化树分析表明,这两个基因均属于DREB1家族,与双子叶植物进化关系较近。实时定量PCR结果显示,CcCBFc和CcCBFd都能被低温(4℃)、干旱(20%PEG)、盐(250 mmol·L~(-1) NaCl)和ABA(100μmol·L~(-1))诱导,表明CcCBFc和CcCBFd可能在香樟应对非生物胁迫过程中发挥重要作用。
Using Cinnamomum camphora seedlings as experimental material, the full-length cDNA sequences of two cold-inducible transcription factor genes CBF / DREB1 were obtained by homologous cloning combined with RACE technology, named as CcCBFc and CcCBFd. The Gen Bank accession numbers were KP336741 and KP336742. Sequence analysis showed that these two genes had no intron. The full-length cDNA was 897 bp and 1 010 bp in length. The open reading frames were 654 bp and 621 bp, encoding 217 and 206 amino acids. The predicted protein molecular weights were 24.0 and 22.9 kD, The isoelectric points were 5.26 and 8.58 respectively. Homology comparison based on amino acid sequence and phylogenetic tree analysis showed that both genes belong to DREB1 family and have close evolutionary relationship with dicotyledonous plants. The results of real-time PCR showed that CcCBFc and CcCBFd could all be treated by low temperature (4 ℃), drought (20% PEG), salt (250 mmol·L -1 NaCl) and ABA (100μmol·L -1) Induced, indicating that CcCBFc and CcCBFd may play an important role in the process of camphor treatment of abiotic stress.