哈萨克羊MSTN基因的DNA甲基化分析
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摘要
【目的】研究肌肉和脂肪组织中MSTN基因(生长抑素)的DNA甲基化水平,为改良哈萨克羊品种的选育提供遗传科学依据。【方法】基于亚硫酸氢盐的方法,测序哈萨克绵羊MSTN基因在Cp G二核苷酸中的DNA甲基化水平。【结论】6月龄哈萨克羊肌肉MSTN基因DNA甲基化低,脂肪组织DNA甲基化程度最高,股二头肌,股三头肌,半膜肌,半腱肌,背最长肌和脂肪组织的MSTN基因的DNA甲基化概率分别为34. 7%,22. 6%,23. 3%,28%,42. 6%和76. 7%。【结论】哈萨克羊各组织MSTN基因DNA的甲基化概率没有显著差异。脂肪组织MSTN基因DNA甲基化的最高,肌肉组织MSTN基因的DNA甲基化的概率较低。脂肪的甲基化概率是背最长肌,股二头肌,股三头肌,半腱肌,半膜肌的甲基化概率分别为:1. 8倍,2. 2倍,3. 39倍,2. 74倍,3. 29倍。其次甲基化从高到低的依次为:脂肪组织>背最长肌>股二头肌>半腱肌>半膜肌>股三头肌。
【Objective】To study the DNA methylation level of MSTN(spermatostatin) gene in muscle and adipose tissue,and to provide genetic data for improving the breeding of Kazakh sheep.【Method】The bisulfite treatment sequencing method using whole genome methylation analysis was used to determine the DNA methylation level of MSTN gene of Kazak sheep in CpG dinucleotide acid. 【Result】The MSTN gene DNA methylation was low in 6-month-old Kazak sheep muscle,and the highest DNA methylation degree in adipose tissue. The DNA methylation probability of MSTN gene in biceps femoris muscle,triceps femoris muscle,semimembranous muscle,semitendinosus muscle,longissimus dorsi muscle and adipose tissue were as follows:34. 7%,22. 6%,23. 3%,28%,42. 6%,and 76. 7%,respectively.【Conclusion】There is no significant difference in methylation probability of MSTN gene DNA between Kazakh sheep tissues. The methylation probability of fat is the highest,and the methylation probability of muscle tissue is low. The methylation probability of fat is the longest in the back,the methylation probabilities of the femoral second muscle,the trigeminal muscle,the semitendinosus,and the semimembrane muscle are 1. 8,2. 2,3. 38,2. 72,and 3. 27 times,respectively,followed by adipose tissue ≥ longissimusdorsi muscle ≥ femoral muscle ≥ semitendinosus ≥semimembranous muscle ≥ trigeminal muscle.
【Objective】To study the DNA methylation level of MSTN(spermatostatin) gene in muscle and adipose tissue,and to provide genetic data for improving the breeding of Kazakh sheep.【Method】The bisulfite treatment sequencing method using whole genome methylation analysis was used to determine the DNA methylation level of MSTN gene of Kazak sheep in CpG dinucleotide acid. 【Result】The MSTN gene DNA methylation was low in 6-month-old Kazak sheep muscle,and the highest DNA methylation degree in adipose tissue. The DNA methylation probability of MSTN gene in biceps femoris muscle,triceps femoris muscle,semimembranous muscle,semitendinosus muscle,longissimus dorsi muscle and adipose tissue were as follows:34. 7%,22. 6%,23. 3%,28%,42. 6%,and 76. 7%,respectively.【Conclusion】There is no significant difference in methylation probability of MSTN gene DNA between Kazakh sheep tissues. The methylation probability of fat is the highest,and the methylation probability of muscle tissue is low. The methylation probability of fat is the longest in the back,the methylation probabilities of the femoral second muscle,the trigeminal muscle,the semitendinosus,and the semimembrane muscle are 1. 8,2. 2,3. 38,2. 72,and 3. 27 times,respectively,followed by adipose tissue ≥ longissimusdorsi muscle ≥ femoral muscle ≥ semitendinosus ≥semimembranous muscle ≥ trigeminal muscle.
引文
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[4]张恒,何成强,李云龙.哺乳动物DNA甲基化的功能与作用[J].生命的化学,2002,(3):218-220.ZHANG Heng,HE Cheng-qiang,LI Yun-long.(2002).Methylation of DNA Mammalian Function and Action[J]. Life Chemistry,(3):218-220.(in Chinese)
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[6] Lee,S. J.(2002). Suppression of body fat accumulation in myostatin-deficient mice. Journal of Clinical Investigation,109(5):595-601.
[7]杨永生,贺建华,邓惠中,等.肌肉生长抑制素对动物肌肉、脂肪和骨骼的影响[J].动物营养学报,2012,(2):220-225.YANG Yun-sheng,HE Jian-hua,DENG Hui-zhong,et al.(2012). The Effects of Myostatin on Muscle,Fat and Bone in Animals[J]. Acta Nutritionica Sinica,(2):220-225.(in Chinese)
[8]凡时财,张学工. DNA甲基化的生物信息学研究进展[J].生物化学与生物物理进展,2009,(2):143-150.FAN Shi-cai,ZHANG Xue-gong.(2009). Advances in Bioinformatics of DNA Methylation[J]. Advances in Biochemistry and Biophysics,(2):143-150.(in Chinese)
[9]刘庆昌.遗传学[M].北京:科学出版社,2006:326.LIU Qin-chang.(2006). Genetics[M]. Beijing:Science Press:326.(in Chinese)
[10]沈秀,林月霞,徐琪. DNA甲基化及其生物学功能[J].中国畜牧兽医,2012,39(6):83-86.SHEN Xiu,LIN Yue-xia,XU Qi.(2012). DNA methylation and its biological functions[J]. Chinese Journal of Animal Husbandry and Veterinary Medicine,39(6):83-86.(in Chinese)
[11]申文雯,樊超男,夏露露,等.肥胖小鼠DNA甲基化改变以及n-3多不饱和脂肪酸的影响作用[J].中国儿童保健杂志,2012,20(4):333-335.SHEN Wen-wen,FAN Chao-nan,XIA Lu-lu,et al.(2012). DNA methylation changes in obese mice and the effect of n-3 polyunsaturated fatty acids[J]. China Children's Health Journal,20(4):333-335.(in Chinese)
[12]侯林,许晟迪,张柱霞,等. PP1γ基因DNA甲基化在学习记忆中的作用[J].中国实验动物学报,2017,25(2):146-152.HOU Lin,XU,XU Chneg-di,ZHANG Zu-xia,et al.(2017). Important role of PP1γDNA methylation in learning and memory[J]. Chinese Journal of Laboratory Animal Science,25(2):146-152.(in Chinese)
[13]Yuan,X. L.,Zhang,Z.,Li,B.,Gao,N.,Zhang,H.,&Sangild,P. T.,et al.(2017). Genome-wide dna methylation analysis of the porcine hypothalamus-pituitary-ovary axis. Sci Rep,7(1).
[14]姚力丹,祖玲玲,马晓燕,等.新疆巴什拜羊肌肉生长抑制素基因DNA甲基化与mRNA表达量分析[J].中国畜牧兽医,2017,44(10):2 987-2 993.YAO Li-dan,ZU Ling-ling,MA Xiao-yan,et al.(2017).Analysis of DNA methylation and mRNA expression of myostatin gene in Xinjiang Bashbai sheep[J]. Journal of Animal Husbandry and Veterinary Medicine of China,44(10):2,987-2,993.
[15]祖玲玲. MSTN/Smad信号通路基因在哈萨克羔羊上的表达规律研究与相关性分析[D].乌鲁木齐:新疆农业大学硕士论文,2015.ZU Ling-ling.(2015). Expression and correlation analysis of MSTN/Smad signaling pathway genes in Kazakh lambs[D].Master Dissertation. Xinjiang Agricultural University,Urumqi.(in Chinese)
[2]贾琦珍,杨菊清,罗康波,等.特克斯县哈萨克羊品种资源概况[J].新疆畜牧业,2012,(1):23-26.QIA Qi-zhen,YANG Ju-qing,LUO Kan-bo,et al..(2012). A Survey of Kazakh Sheep Breed Resources in Turks[J]. Animal Husbandry of Xinjiang,(1):23-26.(in Chinese)
[3]龙定彪,张克英,陈代文,等.肌肉生长抑制素的研究进展[J].动物营养学报,2007,(S1):498-502.LONG Ding-biao,ZHANG Ke-ying,CHEN Dai-wen,et al.(2007). Research Progress of Muscle Growth Inhibitors[J].Acta Nutrimenta Sinica,(S1):498-502.(in Chinese)
[4]张恒,何成强,李云龙.哺乳动物DNA甲基化的功能与作用[J].生命的化学,2002,(3):218-220.ZHANG Heng,HE Cheng-qiang,LI Yun-long.(2002).Methylation of DNA Mammalian Function and Action[J]. Life Chemistry,(3):218-220.(in Chinese)
[5]岳志刚,杨福合,邢秀梅,等.肌肉生长抑制素的研究新进展[J].黑龙江畜牧兽医,2010,(11):31-33.YUE Zhi-gang,YAN Fu-he,XING Xiu-mei,et al.(2010). Progress in the Study of Muscle Growth Inhibitors[J].Heilongjiang Animal and Veterinary Medicine,(11):31-33.(in Chinese)
[6] Lee,S. J.(2002). Suppression of body fat accumulation in myostatin-deficient mice. Journal of Clinical Investigation,109(5):595-601.
[7]杨永生,贺建华,邓惠中,等.肌肉生长抑制素对动物肌肉、脂肪和骨骼的影响[J].动物营养学报,2012,(2):220-225.YANG Yun-sheng,HE Jian-hua,DENG Hui-zhong,et al.(2012). The Effects of Myostatin on Muscle,Fat and Bone in Animals[J]. Acta Nutritionica Sinica,(2):220-225.(in Chinese)
[8]凡时财,张学工. DNA甲基化的生物信息学研究进展[J].生物化学与生物物理进展,2009,(2):143-150.FAN Shi-cai,ZHANG Xue-gong.(2009). Advances in Bioinformatics of DNA Methylation[J]. Advances in Biochemistry and Biophysics,(2):143-150.(in Chinese)
[9]刘庆昌.遗传学[M].北京:科学出版社,2006:326.LIU Qin-chang.(2006). Genetics[M]. Beijing:Science Press:326.(in Chinese)
[10]沈秀,林月霞,徐琪. DNA甲基化及其生物学功能[J].中国畜牧兽医,2012,39(6):83-86.SHEN Xiu,LIN Yue-xia,XU Qi.(2012). DNA methylation and its biological functions[J]. Chinese Journal of Animal Husbandry and Veterinary Medicine,39(6):83-86.(in Chinese)
[11]申文雯,樊超男,夏露露,等.肥胖小鼠DNA甲基化改变以及n-3多不饱和脂肪酸的影响作用[J].中国儿童保健杂志,2012,20(4):333-335.SHEN Wen-wen,FAN Chao-nan,XIA Lu-lu,et al.(2012). DNA methylation changes in obese mice and the effect of n-3 polyunsaturated fatty acids[J]. China Children's Health Journal,20(4):333-335.(in Chinese)
[12]侯林,许晟迪,张柱霞,等. PP1γ基因DNA甲基化在学习记忆中的作用[J].中国实验动物学报,2017,25(2):146-152.HOU Lin,XU,XU Chneg-di,ZHANG Zu-xia,et al.(2017). Important role of PP1γDNA methylation in learning and memory[J]. Chinese Journal of Laboratory Animal Science,25(2):146-152.(in Chinese)
[13]Yuan,X. L.,Zhang,Z.,Li,B.,Gao,N.,Zhang,H.,&Sangild,P. T.,et al.(2017). Genome-wide dna methylation analysis of the porcine hypothalamus-pituitary-ovary axis. Sci Rep,7(1).
[14]姚力丹,祖玲玲,马晓燕,等.新疆巴什拜羊肌肉生长抑制素基因DNA甲基化与mRNA表达量分析[J].中国畜牧兽医,2017,44(10):2 987-2 993.YAO Li-dan,ZU Ling-ling,MA Xiao-yan,et al.(2017).Analysis of DNA methylation and mRNA expression of myostatin gene in Xinjiang Bashbai sheep[J]. Journal of Animal Husbandry and Veterinary Medicine of China,44(10):2,987-2,993.
[15]祖玲玲. MSTN/Smad信号通路基因在哈萨克羔羊上的表达规律研究与相关性分析[D].乌鲁木齐:新疆农业大学硕士论文,2015.ZU Ling-ling.(2015). Expression and correlation analysis of MSTN/Smad signaling pathway genes in Kazakh lambs[D].Master Dissertation. Xinjiang Agricultural University,Urumqi.(in Chinese)