Characterization of protein-DNA interactions using protein microarrays.
Characterization of protein-DNA interactions using protein microarrays.
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DOI：
10.1101/pdb.prot5614
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发表时间：
2011-05-01
影响因子：
--
通讯作者：
Zhu H
中科院分区：
文献类型：
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作者：
Hu S;Xie Z;Blackshaw S;Qian J;Zhu H
蛋白质-DNA相互作用（PDIs）对于许多细胞过程是至关重要的。我们在这里提出了一个协议，在体外使用蛋白质微阵列技术鉴定PDIs。该程序包括用荧光标记的引物将合成的DNA寡核苷酸双链化，将标记的双链DNA直接结合到蛋白质微阵列上，并分析所得PDI的结合。这种方法可以同时识别数千种蛋白质的PDI，并且可以并行测试多个精心设计的DNA探针，这使得能够在蛋白质组范围内快速绘制PDI。
Protein–DNA interactions (PDIs) are critical for many cellular processes. We present here a protocol for the identification of PDIs in vitro using protein microarray technology. The procedure involves doublestranding synthesized DNA oligonucleotides with a fluorescent-labeled primer, binding the labeled double-stranded DNA directly to the protein microarray, and analyzing binding of the resulting PDIs. This approach provides simultaneous identification of PDIs for thousands of proteins, and multiple carefully designed DNA probes can be tested in parallel, which enables a rapid mapping of PDIs on a proteome-wide scale.