ITGB2基因在苏博美利奴羊不同细度皮肤组织中的DNA甲基化及mRNA表达水平分析

doi:10.16431/j.cnki.1671-7236.2020.04.005

Abstract

Abstract: This study was aimed to analyze the DNA methylation and mRNA expression levels of ITGB2 gene in different fine skin tissues of Subo Merino sheep.Using Subo Merino sheep aged ewe as experimental animal and skin tissue samples of different fineness as experimental samples,the promoter CpG island of ITGB2 gene (GenBank accession No.:NC_040252.1) were predicted,the BSP primer,the primers of the mRNA sequence of ITGB2 gene (GenBank accession No.:NM_001009485.1) and GAPDH gene (GenBank accession No.:NM_001190390.1) were designed,respectively.The bisulfite sequencing method (BSP method) was used to amplify and purify,and then inserted into pMD19-T vector and transferred to JM109 cells for overnight culture to form a single colony.The positive clones were screened for sequencing and the obtained sequences were analyzed.The methylation pattern of CpG island in the promoter region of ITGB2 gene in skin tissue of aged ewe,and the mRNA expression level of ITGB2 gene in different fineness skin tissues of Subo Merino sheep was detected by Real-time quantitative PCR.The results showed that the methylation rate of CpG island of extremely fine group in Subo Merino sheep (94.29%) was higher than that of extremely thick group (87.62%).Among them,the methylation rate of CpG2,CpG3,CpG4 and CpG7 island in extremely fine group of Subo Merino sheep (100%,100%,100% and 80.00%) were higher than that in extremely thick group (86.67%,93.33%,80.00% and 73.33%).The expression of ITGB2 gene in extremely thick skin tissues of Subo Merino sheep was extremely significantly higher than that in extremely fine skin tissues(P < 0.01),and there was a significant negative correlation between the level of DNA methylation and the mRNA expression of ITGB2 gene.This results indicated that DNA methylation had a role in skin growth and development,and could be used as a candidate epigenetic marker for Subo Merino sheep.

Key words: Subo Merino sheep; ITGB2 gene; DNA methylation

CLC Number:

S813.3

DU Jianwen, YANG Xuemei, WANG Dan, ZHU Hua, HE Junmin, ABLAT Sulayman, TIAN Yuezhen, ZHAO Bingru, XU Xinming, FU Xuefeng, HANIKEZI Tulafu, WU Weiwei, TIAN Kechuan, HUANG Xixia. Analysis of DNA Methylation and mRNA Expression Levels of ITGB2 Gene in Different Fineness of Subo Merino Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1009-1017.

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Analysis of DNA Methylation and mRNA Expression Levels of ITGB2 Gene in Different Fineness of Subo Merino Sheep

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