Structure and expression of the human gene encoding major heat shock protein HSP70
Structure and expression of the human gene encoding major heat shock protein HSP70
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编码主要热休克蛋白HSP70的人类基因的结构和表达
DOI：
10.1128/mcb.5.2.330-341.1985
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发表时间：
1985
影响因子：
5.3
通讯作者：
R. Morimoto
中科院分区：
文献类型：
--
作者：
Barbara J. Wu;C. Hunt;R. Morimoto
以果蝇热休克蛋白70（HSP 70）基因为异源杂交探针，从人基因组文库中克隆了一个编码70，000-道尔顿热休克蛋白（HSP 70）的基因。人重组克隆与来自暴露于43 ℃ 2小时的HeLa细胞的2.6-腺苷酸酶多聚腺苷酸化mRNA杂交。显示2.6-腺苷酸酶mRNA指导70，000-道尔顿蛋白质的体外翻译，该蛋白质在电泳迁移率方面与体内合成的HSP 70相似。从S1核酸酶抗性的mRNA-DNA杂交分析，HSP 70基因似乎是作为一个不间断的mRNA的2.3酶转录。我们表明，克隆的HSP 70基因包含必要的序列热休克诱导表达的两个标准。首先，用含有HSP 70基因和侧翼序列的亚克隆转染的仓鼠细胞在热休克时合成HSP 70样蛋白。第二，用含有HSP 70基因的5 ′侧翼序列和细菌氯霉素乙酰转移酶基因的编码序列的嵌合基因转染的人细胞在热休克时转录嵌合基因。我们表明，在腺病毒5转化的人细胞系（293细胞）中转录的HSP 70 mRNA与热休克诱导的HSP 70 mRNA相同。
We have cloned a human gene encoding the 70,000-dalton heat shock protein (HSP70) from a human genomic library, using the Drosophila HSP70 gene as a heterologous hybridization probe. The human recombinant clone hybridized to a 2.6-kilobase polyadenylated mRNA from HeLa cells exposed to 43 degrees C for 2 h. The 2.6-kilobase mRNA was shown to direct the translation in vitro of a 70,000-dalton protein similar in electrophoretic mobility to the HSP70 synthesized in vivo. From the analysis of S1 nuclease-resistant mRNA-DNA hybrids, the HSP70 gene appears to be transcribed as an uninterrupted mRNA of 2.3 kilobases. We show that the cloned HSP70 gene contains the sequences necessary for heat shock-induced expression by two criteria. First, hamster cells transfected with a subclone containing the HSP70 gene and flanking sequences synthesized a HSP70-like protein upon heat shock. Second, human cells transfected with a chimeric gene containing the 5' flanking sequences of the HSP70 gene and the coding sequences of the bacterial chloramphenicol acetyltransferase gene transcribed the chimeric gene upon heat shock. We show that the HSP70 mRNA transcribed in an adenovirus 5 transformed human cell line (293 cells) is identical to the HSP70 mRNA induced by heat shock.
影响因子：
5.6
作者：
DEININGER, PL;JOLLY, DJ;SCHMID, CW
通讯作者：
SCHMID, CW