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烟草DREB-like转录因子的克隆与鉴定

Cloning and Identification of DREB-like Transcription Factor in Nicotiana benthamiana

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【作者】 刘卫群王永亮赵同金周海梦郭蔼光

【Author】 LIU Wei-qun1 2, WANG Yong-liang 2, ZHAO Tong-jin3, ZHOU Hai-meng3 , GUO Ai-guang1* (1.College of Life Science, Northwest Agriculture and Forestry University, Yangling 712100, China; 2. Department of Agricultural Biotechnology, Henan Agricultural University, Zhengzhou 450002, China; 3. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China)

【机构】 西北农林科技大学生命科学学院河南农业大学农业生物技术系清华大学生物科学与技术系西北农林科技大学生命科学学院 杨凌712100河南农业大学农业生物技术系郑州450002郑州450002北京100084杨凌712100

【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。

【Abstract】 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.

【关键词】 烟草DREB克隆酵母单杂交反式激活
【Key words】 tobaccoDREBcloneyeast one-hybridtrans-acting
  • 【文献出处】 农业生物技术学报 ,Journal of Agricultural Biotechnology , 编辑部邮箱 ,2006年03期
  • 【分类号】S572
  • 【被引频次】5
  • 【下载频次】337
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