关键词： 3’-UTRs AB ANOVA BSP Bisulfite sequencing PCR CCCTC binding factor COBRA CTCF Cattle Complementary DNA CpG Cyt osineguan inedinucleotide DMR DNA methylation FB H19 IGF2 Insulin-like growth factor 2 LDM QTLs QUMA SEM SPSS TSS TTS adult bovine analysis of variance base pair(s) bp cDNA combined bisulfite restriction analysis differentially methylated region fetal bovine histocompatibility gene longissimus dorsi muscle miRNA microRNA qPCR quantification tool for methylation analysis quantitative real-time PCR quantitative trait locus regulation at the 3’ untranslated regions standard error statistical product and service solutions the transcription termination site transcription start site

Mesh ： Animals Base Sequence Breeding Cattle DNA Methylation Down-Regulation Exons Gene Expression Insulin-Like Growth Factor II / genetics Male Molecular Sequence Data Muscle Development / genetics RNA, Messenger / genetics Tissue Distribution

来  源：   DOI：10.1016/j.gene.2013.09.111   PDF(Sci-hub)

Abstract：
DNA methylation is a key epigenetic modification in mammals and has an essential and important role in muscle development. Insulin-like growth factor 2 (IGF2) is a fetal growth and differentiation factor that plays an important role in muscle growth and in myoblast proliferation and differentiation. The aim of this study was to evaluate the expression of IGF2 and the methylation pattern on the differentially methylated region (DMR) of the last exon of IGF2 in six tissues with two different developmental stages. The DNA methylation pattern was compared using bisulfite sequencing polymerase chain reaction (BSP) and combined bisulfite restriction analysis (COBRA). The quantitative real-time PCR (qPCR) analysis indicated that IGF2 has a broad tissue distribution and the adult bovine group showed significant lower mRNA expression levels than that in the fetal bovine group (P<0.05 or P<0.01). Moreover, the DNA methylation level analysis showed that the adult bovine group exhibited a significantly higher DNA methylation levels than that in the fetal bovine group (P<0.05 or P<0.01). These results indicate that IGF2 expression levels were negatively associated with the methylation status of the IGF2 DMR during the two developmental stages. Our results suggest that the methylation pattern in this DMR may be a useful parameter to investigate as a marker-assisted selection for muscle developmental in beef cattle breeding program and as a model for studies in other species.
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