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首页> 外文期刊>Journal of cellular biochemistry. >Genomic cloning and promoter analysis of the GAHSP40 gene.
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Genomic cloning and promoter analysis of the GAHSP40 gene.

机译：GAHSP40基因的基因组克隆和启动子分析。

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The new heat shock protein (GAHSP40), which binds to Gadd34, is a member of the Hsp40 family gene and has a J domain, which is similar to bacterial DNAJ. We have isolated and sequenced the mouse GAHSP40 gene including 1.6 kb of the 5'-flanking region. Primer extension analysis revealed that the transcription initiation site was located 36-bp upstream of the ATG translation initiation codon. In order to identify the heat-responsive regions in the GAHSP40, NIH3T3 cells were transiently transfected with a series of 5' terminus-truncated mutants of the GAHSP40 promoter linked to the luciferase reporter gene. We found that the region of -284 to -184 bp from initiation start site responded to heat shock treatment. By the gel shift analysis, we found the heat shock elements (HSEs) located in this region from -257 to -225. This HSEs has five 5 bp motifs. The transfection studies using HSEs mutant vectors revealed that those 3' two 5 bp motifs are essential for heat responsive transcription.
机译：与Gadd34结合的新的热激蛋白（GAHSP40）是Hsp40家族基因的成员，具有一个J结构域，与细菌DNAJ相似。我们已经分离并测序了包括1.6 kb的5'侧翼区的小鼠GAHSP40基因。引物延伸分析显示，转录起始位点位于ATG翻译起始密码子上游36 bp。为了鉴定GAHSP40中的热响应区域，用与荧光素酶报道基因连接的一系列GAHSP40启动子的5'末端截短的突变体瞬时转染NIH3T3细胞。我们发现，从起始起始位点开始的-284到-184 bp的区域对热激处理有反应。通过凝胶位移分析，我们发现了从-257到-225的该区域的热激元件（HSE）。该HSE具有五个5bp的基序。使用HSEs突变载体进行的转染研究表明，那些3'两个5 bp的基序对于热响应转录至关重要。

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