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Chinese Journal of Medical Genetics ; (6): 284-288, 2012.
Article in Chinese | WPRIM | ID: wpr-295493

ABSTRACT

<p><b>OBJECTIVE</b>To develop a method for enriching methylated DNA in clinical samples using mesocellular silica foams (MCFs) immobilized with methyl-CpG binding domain (MBD).</p><p><b>METHODS</b>MCFs with ultra-large pore size were synthesized, functionalized and immobilized with GST-MBD.</p><p><b>RESULTS</b>The large cage-like pore structures of MCF materials was retained after functionalization and immobilization, with pore diameter of 55 nm, window size of 30 nm, and a high pore volume of 1.0 cm(3)/g. The loading amount of MBD was as high as 53 wt%. Immobilized MBD showed high binding activity and stability. In a binding buffer with salt concentrations ranging 500-550 mmol/L, the MCF-MBD can selectively enrich methylated DNA from the mixed DNA solution.</p><p><b>CONCLUSION</b>The MCF-MBD method may offer a better choice for high-throughout DNA methylation screening, and has laid a foundation for clinical application, prenatal diagnosis and research on DNA methylation-related genetic diseases.</p>


Subject(s)
Animals , Rats , CpG Islands , DNA , Chemistry , Genetics , Metabolism , DNA Methylation , DNA-Binding Proteins , Chemistry , Immobilized Proteins , Chemistry , Protein Structure, Tertiary , Silicon Dioxide , Chemistry
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