Creation and Function of Novel Gene Regulation Molecules Based on Zinc Finger Motif

On the basis of the DNA recognition mode and the structural features unique to the Cysy D22-His Cysy D22-His Cysy D2 class of nucleic acid binding， approaches to link Cysy D22-His Cysy D22-His Cysy D22-Dinc fingers with other功能模块such as DNA binding domains to generate artificial chemeric peptides with长绑定位点， and DNA-cleavage modules to produce novel sequence specific nucleases，イイD14-5エD1 have been performed。从统计上讲，假定随机基分布，一个独特的16-或18-基对序列将在4.3亿或680亿个核苷酸中只出现一次，并且与人类基因组的大小相同或较大。Although Natural Protein包含长的聚乳酸阵列，如锌手指多-mains从序列中被干扰，没有锌手指蛋白被证明绑定在一起这么长的，连续的DNA序列。因此，这是一个特殊的兴趣，即六指或九指肽被创造和演示在特定序列时尚中结合18或27个连续的碱基对DNA。一种人工蛋白质，它表明DNA修饰性变化是对特定基因的转录调节器。我们创建了6-锌手指蛋白，Sp1 ZF 6（Gly）n（n=4，7，10），通过与柔性聚glycine肽连接器连接两个DNA绑定因子Sp1的转录因子域，并与本机3-锌手指Sp1（530-623）比较DNA结构的影响。新设计的6指肽Sp1 ZF（Gly）7和Sp1 ZF（Gly）10可以在两个分离绑定站点的间隔区域诱导DNA弯曲，而两个3指手指动力在DNA弯曲方向上具有关键效应。这类聚乳酸锌手指肽应该是新的转基因植物和动物的发展。

On the basis of the DNA recognition mode and the structural features unique to the CysィイD22ィエD2-HisィイD22ィエD2 class of nucleic acid binding, approaches to link CysィイD22ィエD2-HisィイD22ィエD2 zinc fingers with other functional modules such as DNA binding domains to generate artificial chimeric peptides with long binding sites, and DNA-cleavage modules to produce novel sequence specific nucleases, ィイD14-5ィエD1 have been performed. Statistically, assuming random base distribution, a unique 16-or 18-base pair sequence will occur only once in 4.3 billion or 68 billion nucleotides, roughly the same or big size of a human genome. Although natural proteins containing long polydactyl arrayl arrays of zinc finger do-mains have been inferred from sequence, no zinc finger proteins have been demonstrated to bind such a long, contiguous DNA sequence. Therefore, it is of special interest that six-or nine-fingered peptide was created and demonstrated to bind 18 or 27 contiguous base pairs of DNA in a sequence specific fashion. An artificial protein that induces a DNA conformational change is interesting as a transcriptional regulator of a specific gene. We created 6-zinc finger proteins, Sp1ZF6(Gly)n(n=4,7,10), by connecting two DNA binding domains of transcription factor Sp1 with flexible polyglycine peptide linkers, and their effects on DNA structure were compared with that of native 3-zinc finger Sp1(530-623). Of special interest are the fact that newly designed 6-finger peptides Sp1ZF(Gly)7 and Sp1ZF(Gly)10 can induce DNA bending at the intervening region of the two distal binding sites and that the liker length between two 3-zinc finger motifs has a crucial effect on the entire DNA bending direction. Such polydactyl zinc finger peptides should be the development of novel transgenic plants and animals.