Gupta R, Webster C I, Gray J C
Department of Plant Sciences and Cambridge Centre for Molecular Recognition, University of Cambridge, UK.
Plant Mol Biol. 1998 Apr;36(6):897-907. doi: 10.1023/a:1005928219895.
The single-copy gene encoding the Arabidopsis HMG-I/Y protein was isolated and characterised. The gene encodes a protein of 204 amino acid residues and contains a single intron of 73 bp. Primer extension analysis indicates that transcription starts 115 bp upstream of the translation start and the leader sequence contains a short open reading frame of 13 amino acid residues. The 5'-upstream region of 2117 bp and several 5' deletions were fused to the beta-glucuronidase (GUS) reporter gene and transferred to tobacco by Agrobacterium-mediated transformation. Analysis of transgenic tobacco plants containing HMG-I/Y promoter regions of -2117, -1468 and -707 from the translation start detected GUS activity in all organs examined, including roots, stems, leaves and floral organs. Deletion from -707 to -185 resulted in a 20-30-fold reduction in GUS activity in roots and stems, indicating the presence of important quantitative regulatory elements in this region.
分离并鉴定了编码拟南芥HMG-I/Y蛋白的单拷贝基因。该基因编码一个由204个氨基酸残基组成的蛋白质,并含有一个73 bp的单一内含子。引物延伸分析表明转录起始于翻译起始位点上游115 bp处,前导序列包含一个由13个氨基酸残基组成的短开放阅读框。将2117 bp的5'-上游区域和几个5'缺失片段与β-葡萄糖醛酸酶(GUS)报告基因融合,并通过农杆菌介导的转化转移到烟草中。对含有从翻译起始位点起-2117、-1468和-707的HMG-I/Y启动子区域的转基因烟草植株进行分析,在所有检测的器官中均检测到GUS活性,包括根、茎、叶和花器官。从-707缺失到-185导致根和茎中GUS活性降低20-30倍,表明该区域存在重要的定量调控元件。