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首页> 外文期刊>FEBS Letters >Identification of human GC‐box‐binding zinc finger protein, a new Krüppel‐like zinc finger protein, by the yeast one‐hybrid screening with a GC‐rich target sequence
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Identification of human GC‐box‐binding zinc finger protein, a new Krüppel‐like zinc finger protein, by the yeast one‐hybrid screening with a GC‐rich target sequence

机译:通过富含GC的靶序列的酵母单杂交筛选鉴定人GC盒结合锌指蛋白,一种新的类似于Krüppel的锌指蛋白。

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摘要

>A new human zinc finger DNA-binding protein was identified by using a yeast one-hybrid selection system. Two versions of the cDNA, encoding the same protein, were detected that differ for a 584 bp extension at the 5′ region. Sequence analysis showed that the longer clone is a full length version containing part of the 5′ untranslated region. The smaller version was fused in frame with the yeast GAL4 activation domain whereas the 5′ region of the longer clone displayed a stop codon interrupting the fusion with the GAL4 domain. Nevertheless, this clone activated the yeast HIS3 reporter gene with the same efficiency as the smaller version. Sequence comparison of the derived protein with the database showed that it belongs to a family of zinc finger DNA-binding proteins which regulate the expression of genes involved in cell proliferation. Expression of the protein in an in vitro system, DNA-binding studies and genetic experiments identify this factor as a new zinc finger DNA-binding protein which binds GC-rich sequences and contains a domain probably functioning as a transcriptional activator. The new human protein identified in this study was therefore named GC-box-binding zinc finger protein).
机译:通过使用酵母单杂交选择系统鉴定了一种新的人锌指DNA结合蛋白。检测到编码相同蛋白质的两个版本的cDNA,在5'区的584 bp延伸不同。序列分析表明,较长的克隆是包含5'非翻译区一部分的全长版本。较小的版本与酵母GAL4激活结构域框内融合,而较长克隆的5'区域显示终止密码子,中断了与GAL4结构域的融合。然而,该克隆以与较小版本相同的效率激活了酵母HIS3报道基因。衍生蛋白与数据库的序列比较表明,它属于锌指DNA结合蛋白家族,可调节参与细胞增殖的基因的表达。该蛋白在体外系统中的表达,DNA结合研究和遗传实验确定该因子为新的锌指DNA结合蛋白,该蛋白与富含GC的序列结合,并包含可能起转录激活因子作用的结构域。因此,在这项研究中鉴定出的新人类蛋白称为GC-box-binding锌指蛋白。

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