Mirault M E, Southgate R, Delwart E
EMBO J. 1982;1(10):1279-85. doi: 10.1002/j.1460-2075.1982.tb00025.x.
Heat-shock genes coding for heat-shock protein 70 (HSP70) in Drosophila melanogaster were subcloned into an SV40/plasmid recombinant capable of replication in permissive monkey COS cells. Following transfection of COS cells, no significant amount of Drosophila hsp70 RNA was detected at 37 degrees C. In contrast, a heat-shock at 43 degrees C or arsenite poisoning at 37 degrees C induced the massive production of Drosophila hsp70 RNA of correct size and faithful 5' ends. After heat-shock, the efficiency of hsp70 transcription in COS cells containing 2-4 X 10(4) gene copies was found to be 15-30% of that measured in Drosophila, on a per gene basis. By testing a series of 5' deletion mutants in this inducible transcription assay it was found that a sequence less than 70 bp long, directly upstream of the hsp70 gene, was essential for the heat or arsenite induction of transcription.
将果蝇中编码热休克蛋白70(HSP70)的热休克基因亚克隆到一种能够在允许的猴COS细胞中复制的SV40/质粒重组体中。转染COS细胞后,在37℃未检测到大量果蝇hsp70 RNA。相反,43℃的热休克或37℃的亚砷酸盐中毒诱导产生了正确大小且5'端忠实的大量果蝇hsp70 RNA。热休克后,发现含有2 - 4×10⁴个基因拷贝的COS细胞中hsp70转录效率,以每个基因计算,是在果蝇中测得的15% - 30%。通过在这种可诱导转录试验中测试一系列5'缺失突变体,发现hsp70基因直接上游一个长度小于70 bp的序列对于热或亚砷酸盐诱导转录至关重要。