Development of a method for methylated DNA enrichment with functionalized mesocellular silica foams immobilized with methyl CpG binding domain / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
; (6): 284-288, 2012.
Article
in Zh
| WPRIM
| ID: wpr-295493
Responsible library:
WPRO
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>
Full text:
1
Database:
WPRIM
Main subject:
DNA
/
Chemistry
/
Protein Structure, Tertiary
/
Silicon Dioxide
/
CpG Islands
/
DNA Methylation
/
DNA-Binding Proteins
/
Immobilized Proteins
/
Genetics
/
Metabolism
Limits:
Animals
Language:
Zh
Journal:
Chinese Journal of Medical Genetics
Year:
2012
Document type:
Article