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首页> 外文期刊>Animals >Cloning and Expression Analysis of Two Kdm Lysine Demethylases in the Testes of Mature Yaks and Their Sterile Hybrids
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Cloning and Expression Analysis of Two Kdm Lysine Demethylases in the Testes of Mature Yaks and Their Sterile Hybrids

机译：两次KDM赖氨酸去甲基酶在成熟牦牛睾丸中的克隆及表达分析及其无菌杂种

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The objective of this study was to explore the molecular mechanism for male sterility of yak hybrids based on two demethylases. Total RNA was extracted from the testes of adult yaks ( n = 10) and yak hybrids (cattle–yaks, n = 10). The coding sequences (CDS) of two lysine demethylases (KDMs), KDM1A and KDM4B, were cloned by RT-PCR. The levels of KDM1A and KDM4B in yaks and cattle–yaks testes were detected using Real-time PCR and Western blotting for mRNA and protein, respectively. In addition, the histone methylation modifications of H3K36me3 and H3K27me3 were compared between testes of yaks and cattle–yaks using ELISA. The CDS of KDM1A and KDM4B were obtained from yak testes. The results showed that the CDS of KDM1A exhibited two variants: variant 1 has a CDS of 2622 bp, encoding 873 amino acids, while variant 2 has a CDS of 2562 bp, encoding 853 amino acids. The CDS of the KDM4B gene was 3351 bp in length, encoding 1116 amino acids. The mRNA and protein expression of KDM1A and KDM4B, as well as the level of H3K36me3, were dramatically decreased in the testes of cattle–yaks compared with yaks. The present results suggest that the male sterility of cattle–yaks might be associated with reduced histone methylation modifications.
机译：本研究的目的是探讨基于两种去甲基酶的牦牛杂交种的雄性不育的分子机制。从成年牦牛（n = 10）和牦牛杂交种（牛牦牛，n = 10）中提取总RNA。通过RT-PCR克隆两个赖氨酸脱乙基酶（KDMS），KDM1A和KDM4B的编码序列（CDS）。使用实时PCR和蛋白质印迹分别检测牦牛和牛牦牛睾丸中KDM1A和KDM4B的水平，分别用于mRNA和蛋白质。此外，使用ELISA比较了牦牛和牛牦牛睾丸的组蛋白甲基化修饰和H3K27ME3。 KDM1A和KDM4B的CD是从牦牛睾丸获得的。结果表明，KDM1A的CD表现出两种变体：变体1具有2622bp的Cd，编码873个氨基酸，而变体2具有2562bp的Cd，编码853氨基酸。 KDM4B基因的CD长度为3351bp，编码1116个氨基酸。与牦牛相比，KDM1A和KDM4B的mRNA和蛋白表达以及H3K36ME3的水平在牛牦牛的睾丸中显着降低。本结果表明，牛牦牛的雄性不育可能与减少的组蛋白甲基化修饰有关。

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