Development of a method for methylated DNA enrichment with functionalized mesocellular silica foams immobilized with methyl CpG binding domain.
- VernacularTitle:应用功能化氧化硅泡沫负载甲基化结合结构域建立一种富集甲基化DNA的方法
- Author:
Ya-ting CHEN
1
;
Lu HAN
;
Dong-yuan ZHAO
;
Bo TU
;
Duan MA
Author Information
- Publication Type:Journal Article
- MeSH: Animals; CpG Islands; DNA; chemistry; genetics; metabolism; DNA Methylation; DNA-Binding Proteins; chemistry; Immobilized Proteins; chemistry; Protein Structure, Tertiary; Rats; Silicon Dioxide; chemistry
- From: Chinese Journal of Medical Genetics 2012;29(3):284-288
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop a method for enriching methylated DNA in clinical samples using mesocellular silica foams (MCFs) immobilized with methyl-CpG binding domain (MBD).
METHODSMCFs with ultra-large pore size were synthesized, functionalized and immobilized with GST-MBD.
RESULTSThe 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.
CONCLUSIONThe 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.