牛Ascl2 基因印记及DNA甲基化状态分析

doi:10.11843/j.issn.0366-6964.2014.12.006

Abstract

Abstract:

In order to determine the imprinting status of Ascl2 in bovines different tissues，and to reveal the possible role of DNA methylation in regulating Ascl2 gene imprinting，we analyzed the expression of Ascl2 in 7 organs (heart，liver，spleen，lung，kidney，muscle and subcutaneous fat) of calves.Single nucleotide polymorphisms were used to identify imprinting by direct sequencing the RT-PCR products，then the DNA methylation status in promoter region of Ascl2 gene were analyzed in lung，liver and kidey tissues of cattle using bisulfite sequencing analysis.The results of RT-PCR showed that Ascl2 gene monoallelic expressed in lung and liver tissues while biallelic expressed in heart，spleen，kidney，muscle and subcutaneous fat tissues.The results of bisulfite sequencing showed that the methylation level of 2 parental strands exhibited extremely significant difference (P<0.01) in lung and liver tissues where Ascl2 was monoallelic expression.The mean methylation levels of A-strands (61.21%，87.28%) showed seriously hypermethylated compared with G-strands (24.70%，25.61%).However，in kidey tissues where Ascl2 was biallelic expression，A-strand (54.70%) and G-strand (49.55%) had no significant difference（P>0.05）.Our results suggest that the differential methylation of CpG island in promoter of Ascl2 regulate the imprinting expression of Ascl2 in lung and liver tissues.

Key words: Ascl2, imprinting status, DNA methylation, differential methylation

CLC Number:

S823
S813.3

WANG Meng-nan，CUI Ya-li,WU Guo-jiang，LI Dong-jie,LI Shi-jie. Genomic Imprinting and DNA Methylation Status of Ascl2 Gene in Cattle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(12): 1949-1956.

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Genomic Imprinting and DNA Methylation Status of Ascl2 Gene in Cattle

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