| 摘要: |
| 【目的】克隆银中杨抗逆转录因子DREB基因，为其功能的深入研究奠定基础。【方法】根据DREB AP2/EREBP保守区设计1对简并引物，采用PCR方法对银中杨（Populus albaxp）转录因子DREB基因中间片段进行克隆；再利用反向PCR方法对该基因的旁侧序列进行克隆，最终将两侧序列与中间片段拼接得到基因全长；同时，根据基因序-列设计1对特异引物，利用荧光定量PCR分析银中杨DREB基因在不同逆境胁迫中的表达情况。【结果】成功地从银中杨中克隆到1个DREB基因，命名为PaDREB（注册号：EF582843）。该基因序列长935 bp，ORF为819 bp，编码272个氨基酸；同源性比较与系统发育分析证实，银中杨DREB属于DREB转录因子家族，并与月季DREB2C具有较高的同源性；荧光定量PCR检测结果显示，盐、低温、干旱及ABA均能诱导DREB基因的表达。【结论】首次从银中杨中分离并克隆了DREB基因，并证实其参与了银中杨对盐、低温、干旱及ABA的应答。 |
| 关键词: 银中杨 DREB转录因子 基因克隆 基因表达 |
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| 基金项目:国家自然科学基金项目（30971804）；国家转基因生物新品种培育重大专项“基因表达调控技术研究”（2010ZX08010-002）；教育部新世纪优秀人才计划项目（NCET-08-0693）；长春市科技局计划项目（09YJ24） |
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| Cloning and expression of DREB gene from Populus albaxp |
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| Abstract: |
| 【Objective】The study was done to clone the stress induced DREB transcription factor gene from Populus albaxp and provide a starting point for the further studies in functional analysis.【Method】We firstly cloned the partial fragment of DREB gene from Populus albaxp using homology cloning methods,according to AP2/EREBP conserved sequences.Then,the flanking sequence of DREB gene was obtained using reverse PCR.The flanking sequences and the partial fragment were combined to a full length sequence,named PaDREB.A real-time PCR was performed to observe the expression profiling of PaDREB under various stresses using a pair of specific primers.【Result】We obtained the full length of PaDREB gene and submitted to Genbank (Accessed Number:EF582843).The full-length of cDNA is 935 bp and the open reading frame (ORF) is 819 bp,encoding 272 amino acids;Phylogenetic trees showed that PaDREB belongs to the DREB super family and shared a higher identity with the DREB2C.According to real-time PCR analysis,we found that salt,low temperature,drought and ABA could all induce the expression of PaDREB dramatically.【Conclusion】We first cloned PaDREB gene from Populus albaxp.The results of RT-PCR showed that PaDREB gene could be involved in the response of salt,low temperature,drought and ABA treatment. |
| Key words: Populus albaxp DREB transcription factor gene cloning gene expression |
