Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus of D. melanogaster.
Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus of D. melanogaster.
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hsp83 转录物在来自黑腹果蝇 63B 热休克基因座的 3292 个核苷酸序列内的定位。
DOI：
10.1093/nar/11.20.7011
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发表时间：
1983
影响因子：
14.9
通讯作者：
Lis,JT
中科院分区：
文献类型：
--
作者：
Hackett,RW;Lis,JT
本研究测定了来自D.黑腹果蝇在此片段中，我们通过S1作图和从限制性片段引物mRNA合成cDNA，定位了编码83，000 d热休克多肽的独特基因（hsp 83基因）的转录起始位点和RNA剪接位点。该序列起始于转录起始点上游879 bp处，包括149 bp非翻译第一外显子、1139 bp内含子，并延伸1125 bp进入蛋白质编码区。这些数据确定了83，000 d多肽的前375个氨基酸的单个开放翻译阅读框架，从位于3′内含子-外显子连接处的第一个ATG密码子开始。我们讨论并演示了E. coilexonuclease III产生单链DNA探针作为替代链分离的mRNA的S1定位。我们还使用同源性搜索标准的基础上已知的蛋白质-DNA结合位点比较我们的热休克蛋白83序列与其他sequencedDrosophila热休克基因。结果表明，hsp 83基因转录起始点附近约80 bp的区域与hsp 70基因的相应区域的同源性仅为31%，而hsp 22、23、26和27基因在该区域与hsp 70的同源性为54%。热休克蛋白83基因的同源性较低，这与热休克基因的异常性质一致。
We have determined the complete nucleotide sequence of a 3292 bp cloned segment derived from the 63B heat shock cytogenetic locus ofD. melanogaster. Within this segment we have positioned the start of transcription and RNA splice sites of the unique gene that encodes the 83,000 d heat shock polypeptide (hsp83 gene) by S1 mapping and synthesis of cDNA from restriction fragment primed mRNA. The sequence begins at a point 879 bp upstream from the transcription start and includes the 149 bp nontranslated first exon, the 1139 bp intron and extends 1125 bp into the protein coding region. These data identify a single open translation reading frame for the first 375 amino acids of the 83,000 d polypeptide, beginning with the first ATG codon located at the 3′ intron-exon junction. We discuss and demonstrate the use ofE. coilexonuclease III generated single-strand DNA probes as an alternative to strand separation for S1 mapping of mRNA. We also use homology search criteria based upon known protein-DNA binding sites to compare our hsp83 sequence with other sequencedDrosophilaheat shock genes. These comparisons indicate that a large region of approximately 80 bp centered around the transcription initiation point of the hsp83 gene shares only a 31% homology with the corresponding region of the hsp70 gene, whereas the hsp22, 23, 26, and 27 genes 8hare a 54% homology with hsp70 in this region. The lower homology of the hsp83 gene is consistent with the deviant nature of this heat shock gene.