Shoji T, Kato K, Sekine M, Yoshida K, Shinmyo A
Metabolic Regulation in Plant Cells, Department of Biological Sciences Nara Institute of Science and Technology (NAIST), 8916-5 Takayama-cho, Ikoma-shi, Nara 630-0101, Japan e-mail:
Plant Cell Rep. 2000 Mar;19(4):414-420. doi: 10.1007/s002990050749.
We have isolated two types of complementary DNA (cDNA) sequences showing high similarity with the DNA-binding domain in heat shock factors (HSFs) from a cDNA library of tobacco suspension-cultured cells (Nicotiana tabacum L. cv. BY2). These two genes, NtHSF1 and NtHSF2, showed low similarity (37%) with each other. Both contained the typical conserved regions of HSFs (DNA-binding domains, leucine zipper repeats, nuclear localization signals, and tryptophan repeats). Transcripts of NtHSF1 and NtHSF2 were detected even at the normal temperature. The recombinant NtHSF1 and NtHSF2 proteins expressed in Escherichia coli bound specifically to a 183-bp DNA fragment of the Arabidopsis thaliana HSP18.2 promoter containing three sets of head-to-head and tail-to-tail repeats of the conserved pentamer unit (nGAAnnTTCnnGAAn).
我们从烟草悬浮培养细胞(烟草品种BY2)的cDNA文库中分离出两种互补DNA(cDNA)序列,它们与热激因子(HSF)中的DNA结合结构域具有高度相似性。这两个基因,NtHSF1和NtHSF2,彼此间的相似性较低(37%)。二者均包含HSF的典型保守区域(DNA结合结构域、亮氨酸拉链重复序列、核定位信号和色氨酸重复序列)。即使在常温下也能检测到NtHSF1和NtHSF2的转录本。在大肠杆菌中表达的重组NtHSF1和NtHSF2蛋白与拟南芥HSP18.2启动子的一个183 bp DNA片段特异性结合,该片段包含三组保守五聚体单元(nGAAnnTTCnnGAAn)的头对头和尾对尾重复序列。