香猪卵巢RARRES1基因第1外显子CG位点的甲基化及其与基因差异表达的关联

doi:10.16431/j.cnki.1671-7236.2018.06.006

Abstract

Abstract:

From the transcriptome data of ovary using the RNA-seq technology, retinoic acid receptor responder 1 (RARRES1) gene was screened out from the differential expression profile of Xiang pigs.To further reveal the underlying reason for the gene differential expression,the ovarian tissues samples were collected from two groups of Xiang pig with high and low litter sizes.Based on bioinformatics analysis which predicted that the downstream of ATG start codon in exon 1 of RARRES1 gene was rich with CpG sites,specific primers for this region were designed,and the methylation status of RARRES1 gene was tested by bisulfite sequencing PCR method (BSP).The expression level of RARRES1 gene was detected by quantitative Real-time PCR method,and its relationship with methylation level was analyzed.The results showed that the methylation level of RARRES1 gene in high litter size group was higher,and in both of the two groups, there were four CpG sites (CpG_7,CpG_11,CpG_12 and CpG_15) keeping unmethylated state,while the other three sites (CpG_8,CpG_17 and CpG_18) were all modified by methylation.There were four CpG sites were differently modified between the two groups with sites,CpG_4(P>0.05),CpG_9(P<0.05) and CpG_16(P<0.05) were higher and site CpG_6 (P<0.01) to be lower in high litter group than that in low litter size group.And the results of quantitative Real-time PCR showed that the mRNA levels of RARRES1 gene in high litter size group were much higher than that in another group (P<0.05).Besides,Spearman correlation analysis estimated there was a strong positive relationship between the methylation ratio of CpG_9 and CpG_16 and the expression level of RARRES1 gene (R²=0.896,P<0.01).It suggested that the methylation of exon 1 region might have a certain effect on promoting the expression of RARRES1 gene in Xiang pig ovary.

Key words: Xiang pig; ovary; RARRES1 gene; methylation; expression level

CLC Number:

S828

RUAN Yiqi, LI Dapeng, RAN Xueqin, MAO Ning, YI Fanli, ZHANG Fuping, WANG Jiafu. The Methylation in the Exon 1 of RARRES1 Gene and its Association with Gene Differential Expression of Xiang Pig Ovary[J]. , 2018, 45(6): 1471-1478.

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The Methylation in the Exon 1 of RARRES1 Gene and its Association with Gene Differential Expression of Xiang Pig Ovary

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