b-Boule基因5′调控序列的克隆与睾丸组织DMR甲基化分析

doi:10.3864/j.issn.0578-1752.2011.18.017

Abstract

Abstract: 【Objective】 In order to reveal b-Boule gene expression and regulation, and to supply evidence for apparent genetic mechanism of cattle-yak male sterile, the 5′-flanking regulatory sequence of b-Boule, and the difference of b-Boule gene DMR methylation status in testes among cattle, yaks and cattle-yaks were studied. 【Method】 The 5′-flanking regulatory sequence of b-Boule in yak was cloned and sequenced by the PCR amplification, the 5´-flanking regulatory sequence of b-Boule was analyzed using bioinformatics, and the DMR methylation status of b-Boule gene in testes among cattle, yaks and cattle-yaks were detected using sodium bisulfite sequencing. 【Result】 The 5′-flanking regulatory sequence length of b-Boule is 1 352 bp, and there is a CpG island. The core promoter region contains SP1 methylation-sensitive site. The methylation level of b-Boule gene DMR in cattle-yaks (17.78%) was significantly greater than that in cattle (6.94%) and yaks (7.50%) (P＜0.01). The difference of the methylation level is obviously, especially the CpG site’s 33-35. 【Conclusion】 The methylation level of yak b-Boule gene DMR is significantly greater than that of cattle-yak. Combined with the previous study results about mRNA expression and histological observation, it was speculated that the DMR methylation played a key role in the expression and regulation of b-Boule, and it might arrest meiotic process of spermatogenesis by its mRNA expression inhibition through the high methylation level of b-Boule gene DMR in cattle-yak.

Key words: yak, cattle-yak, b-Boule gene, 5′regulation sequence, methylation

LI Ming-Gui, XU Hong-Tao, LI Yin-Xia, YU Sha-Li, ZHAO Xing-Bo, PAN Zeng-Xiang, ZHU Xiang, XIE Zhuang, LI Qi-Fa. Cloning and Testicular DMR Methylation Analysis of b-Boule Gene 5′-Flanking Regulation Regions[J].Scientia Agricultura Sinica, 2011, 44(18): 3859-3867.

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Cloning and Testicular DMR Methylation Analysis of b-Boule Gene 5′-Flanking Regulation Regions

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