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首页> 外文期刊>Molecular and Cellular Biology >Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene.
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Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene.

机译：人c-myc基因双启动子内顺式作用调控的靶序列。

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Recombinant plasmids of the human c-myc promoter-leader region and the bacterial chloramphenicol acetyltransferase (cat) gene were constructed. After transfection into different rodent and human cells, the 862-base-pair (bp) PvuII fragment carrying both c-myc promoters and 350 bp of the untranslated leader conferred 1/15 to 1/30 of the CAT activity mediated by the simian virus 40 promoter. The presence of additional sequences upstream of the PvuII fragment had an overall negative effect on c-myc promoter activity detectable by titration analysis with small amounts of transfected plasmid DNA. The analysis of numerous deletion constructs in the c-myc promoter-leader region as well as S1 mapping experiments demonstrated that the high CAT activity depended largely on the presence of the second promoter. By cotransfection of c-myc-cat constructs with plasmids carrying different parts of the c-myc promoter locus, targets for positively acting cellular factors were identified. Two positive regulatory elements were mapped within the 862-bp PvuII fragment. One was localized within the 248-bp PvuII-SmaI fragment -101 to -349 bp upstream of the first cap site and the other within the 142-pb XhoI-NaeI fragment of the first exon, comprising positions -95 to +47 relative to the second cap site. We conclude that the dual promotor of the human c-myc gene represents a strong eucaryotic promotor regulated by cooperation of positively and negatively acting cellular transcription factors.
机译：构建了人c-myc启动子-前导区和细菌氯霉素乙酰转移酶（cat）基因的重组质粒。转染到不同的啮齿动物和人类细胞后，带有c-myc启动子和350 bp未翻译前导序列的862个碱基对（bp）的PvuII片段赋予猿猴病毒介导的CAT活性的1/15至1/30 40个启动子。通过用少量转染质粒DNA进行滴定分析可检测到PvuII片段上游其他序列的存在，对c-myc启动子活性具有总体负面影响。对c-myc启动子-前导区中许多缺失构建体的分析以及S1作图实验表明，高CAT活性很大程度上取决于第二个启动子的存在。通过将c-myc-cat构建体与携带c-myc启动子基因座不同部分的质粒共转染，可以鉴定出具有积极作用的细胞因子的靶标。在862 bp PvuII片段内定位了两个阳性调控元件。一个位于第一个帽位点上游的248-bp PvuII-SmaI片段-101至-349 bp内，另一个位于第一个外显子的142-pb XhoI-NaeI片段内，相对于-95至+47位第二个网站上限。我们得出的结论是，人类c-myc基因的双重启动子代表了一个强的真核启动子，受到正负作用的细胞转录因子的共同调控。

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