Park S M, Hong C B
Institute for Molecular Biology and Genetics, Seoul National University, Korea.
Mol Cells. 1998 Oct 31;8(5):594-9.
We isolated a cDNA clone, named TLHS-1, for a low molecular weight heat-shock protein (LMW HSP) from tobacco. The nucleotide sequence determination of the clone identified an open reading frame for 159 amino acids. To the upstream of the open reading frame, a sequence of 124 nucleotides was determined. To the 3' downstream of the open reading frame, 212 nucleotides were identified which carried a poly(A) tail. A comparison of the open reading frame of TLHS-1 with the previously reported class I LMW HSPs showed a high identity which classified the TLHS-1 as a class I LMW HSP cDNA clone. Reexamination of the amino acid sequences for LMW HSPs including the TLHS-1 argued for the necessity to redraw the consensus region. Six consensus regions were proposed instead of the two consensus regions suggested previously. A RNA blot hybridization for TLHS-1 showed the typical expression pattern of a heat-shock inducible gene from 3 common tobacco cultivars. The genomic DNA blot hybridization for the TLHS-1 gene from the 3 tobacco cultivars showed identical patterns. These results implied that the expression control and structure of the TLHS-1 gene have been well conserved in the tobacco cultivars which are not bred for high temperature stress resistance.
我们从烟草中分离出一个低分子量热休克蛋白(LMW HSP)的cDNA克隆,命名为TLHS-1。对该克隆进行核苷酸序列测定,确定了一个编码159个氨基酸的开放阅读框。在开放阅读框的上游,确定了一段124个核苷酸的序列。在开放阅读框的3'下游,鉴定出212个核苷酸,其带有一个poly(A)尾巴。将TLHS-1的开放阅读框与先前报道的I类LMW HSPs进行比较,显示出高度的同一性,这将TLHS-1归类为I类LMW HSP cDNA克隆。对包括TLHS-1在内的LMW HSPs的氨基酸序列进行重新检查,表明有必要重新绘制共有区域。提出了六个共有区域,而不是先前建议的两个共有区域。对TLHS-1进行的RNA印迹杂交显示了来自3个普通烟草品种的热休克诱导基因的典型表达模式。对这3个烟草品种的TLHS-1基因进行的基因组DNA印迹杂交显示出相同的模式。这些结果表明,TLHS-1基因的表达调控和结构在未经过高温胁迫抗性培育的烟草品种中得到了很好的保守。