Scientific papers // Epigenetics // Real time GlaI-PCR assay of regulation regions of human genes HDAC4, RARB and URB1
Real time GlaI-PCR assay of regulation regions of human genes HDAC4, RARB and URB1
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, AG Akishev, MA Abdurashitov, SB Oleynikova, VL Sitko, and S Kh Degtyarev
RJPBCS, vol 7(2), 2016, p.p. 667-676
Real-time GlaI-PCR assay is developed to determine DNA methylation status of the regulation regions of HDAC4, URB1 and RARB genes in DNA preparations from human leukocytes. Real-time GlaI-PCR assay is DNA hydrolysis with methyl-directed site-specific DNA endonuclese GlaI followed by real-time PCR from the primers located upstream and downstream of the studied DNA region. The obtained data confirm a full methylation of the studied ACGT and GCGC sites in the regulatory regions of the HDAC4 and URB1 genes and a complete hydrolysis of these sites with GlaI. A first exon of RARB gene is slightly methylated in the leukocytes DNA preparations and according to the results of GlaI-PCR assay we don’t observe GlaI hydrolysis of ACGCG site in RARB gene. The data obtained correspond to the literature data. The proposed method of real-time GlaI-PCR assay may be used to determine the methylation status of any unique parts in human and mammalian genomes.