Bulyk Martha L
Division of Genetics, Department of Medicine, Harvard/MIT Division of Health Sciences and Technology (HST), Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Curr Opin Biotechnol. 2006 Aug;17(4):422-30. doi: 10.1016/j.copbio.2006.06.015. Epub 2006 Jul 12.
DNA-binding proteins have key roles in many cellular processes, including transcriptional regulation and replication. Microarray-based technologies permit the high-throughput identification of binding sites and enable the functional roles of these binding proteins to be elucidated. In particular, microarray readout either of chromatin immunoprecipitated DNA-bound proteins (ChIP-chip) or of DNA adenine methyltransferase fusion proteins (DamID) enables the identification of in vivo genomic target sites of proteins. A complementary approach to analyse the in vitro binding of proteins directly to double-stranded DNA microarrays (protein binding microarrays; PBMs), permits rapid characterization of their DNA binding site sequence specificities. Recent advances in DNA microarray synthesis technologies have facilitated the definition of DNA-binding sites at much higher resolution and coverage, and advances in these and emerging technologies will further increase the efficiencies of these exciting new approaches.
DNA结合蛋白在许多细胞过程中发挥关键作用,包括转录调控和复制。基于微阵列的技术允许高通量鉴定结合位点,并能够阐明这些结合蛋白的功能作用。特别是,对染色质免疫沉淀的DNA结合蛋白(ChIP芯片)或DNA腺嘌呤甲基转移酶融合蛋白(DamID)进行微阵列读出,能够鉴定蛋白质在体内的基因组靶位点。一种用于直接分析蛋白质与双链DNA微阵列体外结合的互补方法(蛋白质结合微阵列;PBM),可以快速表征其DNA结合位点序列特异性。DNA微阵列合成技术的最新进展促进了以更高分辨率和覆盖率定义DNA结合位点,这些技术以及新兴技术的进步将进一步提高这些令人兴奋的新方法的效率。