| 烟草DREB-like转录因子的克隆与鉴定 | |
| 刘卫群 ; 王永亮 ; 赵同金 ; 周海梦 ; 郭蔼光 ; LIU Wei-qun ; WANG Yong-liang ; ZHAO Tong-jin ; ZHOU Hai-meng ; GUO Ai-guang | |
| 2010-06-07 ; 2010-06-07 | |
| 关键词 | 烟草 DREB 克隆 酵母单杂交 反式激活 tobacco DREB clone yeast one-hybrid trans-acting S572 |
| 其他题名 | Cloning and Identification of DREB-like Transcription Factor in Nicotiana benthamiana |
| 中文摘要 | 在NCBI网站BLAST得到5个烟草表达序列标签(EST),据此设计引物,从烟草(Nicotianabenthamiana)品种本塞母氏中分离出2条DREB(dehydration-responsiveelementbindingprotein)-like转录因子基因,分别命名为NbDREB1和NbDREB2。与相关AP2/EREBP类转录因子比对结果显示,NbDREB1和NbDREB2都具有典型的AP2/EREBP转录因子家族EREBP亚族A类AP2区特征,并且都能在大肠杆菌(Escherichiacoli)中表达。酵母单杂交结果显示,都不具有激活功能。连接pGADT7反式激活载体进行酵母单杂交结果显示:NbDREB1能与DRE顺式作用元件结合,NbDREB2则不能,表明NbDREB1为抑制型DREB类转录因子。比较NbDREB1和NbDREB2的AP2区发现:两者在第2位和第49位氨基酸处不同,NbDREB1为Y和K,NbDREB2为N和R。; Five EST (expressed sequence tag) sequences were obtained in NCBI web BLAST according to the AP2 domain sequence of DREB(dehydration-responsive binding protein)1a gene from Arabidopsis. Specific primers were designed based on the EST sequences, and two DREB-like genes were amplified from tobacco (Nicotiana benthamiana) cDNA by PCR, and named NbDREB1 and NbDREB2 respectively and could be expressed in Escherichia coli. Comparing with other dehydration-responsive element binding related protein found that NbDREB1 and NbDREB2 possessed typical structural characteristics of A type of EREBP subgroup in AP2/EDREBP transcription factor family . The yeast one-hybrid (expression vector YepGap) showed that the two genes had no active function. Anti-activator yeast one-hybrid (expression vector pGADT7containning GAL4 domain) assay showed that NbDREB1 could bind to DRE cis-element in yeast, but NbDREB2 could not. The difference of AP2 domain between NbDREB1 and NbDREB2 was only in the 2nd and 49th amino acid, which was Y and K in NbDREB1 and N and R in NbDREB2. |
| 语种 | 中文 ; 中文 |
| 内容类型 | 期刊论文 |
| 源URL | [http://hdl.handle.net/123456789/45714]  |
| 专题 | 清华大学 |
| 推荐引用方式 GB/T 7714 | 刘卫群,王永亮,赵同金,等. 烟草DREB-like转录因子的克隆与鉴定[J],2010, 2010. |
| APA | 刘卫群.,王永亮.,赵同金.,周海梦.,郭蔼光.,...&GUO Ai-guang.(2010).烟草DREB-like转录因子的克隆与鉴定.. |
| MLA | 刘卫群,et al."烟草DREB-like转录因子的克隆与鉴定".(2010). |
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