金川多肋牦牛Hoxa6和Hoxa10基因甲基化与mRNA差异表达研究

doi:10.11843/j.issn.0366-6964.2015.09.007

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (9): 1532-1539.doi: 10.11843/j.issn.0366-6964.2015.09.007

金川多肋牦牛Hoxa6和Hoxa10基因甲基化与mRNA差异表达研究

熊显荣^1#，张雁^1#，兰道亮²，李键^2*，字向东¹，李善蓉³，杨建梅³

(1.西南民族大学生命科学与技术学院，成都 610041；2.西南民族大学青藏高原研究院，成都 610041；3.金川县兽医畜牧局，金川 624100)

收稿日期:2014-11-02 出版日期:2015-09-23 发布日期:2015-09-23
通讯作者: 李键，教授，博士生导师，主要从事牦牛细胞生物学和发育生物学研究，E-mail：jianli_1967@163.com
作者简介:熊显荣（1984-），男，江西赣州人，硕士，主要从事牦牛细胞生物学和发育生物学研究，E-mail：xianrongxiong@163.com；张雁（1990-），女，吉林长春人，硕士，主要从事牦牛遗传育种研究，E-mail：150115439@qq.com。二者共同并列为第一作者
基金资助:
国家科技支撑计划（2012BAD13B06）；四川省科技支撑计划（2014NZ0114）；西南民族大学牦牛创新团队资助(13CXTD01)

Differences of Hoxa6 and Hoxa10 Methylation Status and mRNA Expression in Multi-costa Properties of Jinchuan Yak

XIONG Xian-rong^1# ，ZHANG Yan^1# ，LAN Dao-liang²，LI Jian ^2*，ZI Xiang-dong¹，LI Shan-rong³，YANG Jian-mei³

(1.College of Life Science and Technology，Southwest University for Nationalities，Chengdu 610041，China；2.Institute of Qinghai-Tibetan Plateau，Southwest University for Nationalities，Chengdu 610041，China；3.Animal Husbandry and Veterinarian Bureau of Jinchuan，Jinchuan 624100，China)

Received:2014-11-02 Online:2015-09-23 Published:2015-09-23

摘要/Abstract

摘要：

本研究旨在通过检测金川多肋牦牛与普通牦牛中Hoxa6和Hoxa10基因的转录水平和启动子区甲基化状态，为揭示Hox基因在金川多肋牦牛多1对肋骨性状形成中的转录调控机制奠定基础。通过实时荧光定量PCR技术检测Hoxa6和Hoxa10基因在多肋牦牛和普通牦牛中的mRNA表达水平，同时采用重亚硫酸盐测序PCR（Bisulfite-sequencing PCR，BSP）法对Hoxa6和Hoxa10启动子区进行甲基化修饰与克隆测序，分析甲基化状态。结果显示，多肋牦牛中Hoxa6基因表达量显著高于普通牦牛(P<0.05)，而Hoxa10基因表达量显著低于普通牦牛(P<0.05)。Hoxa6启动子区域的CpG2，在普通牦牛中的DNA甲基化显著高于多肋牦牛(P<0.05)，尤其是第3、4、8、20和21位CpG位点；Hoxa10启动子区域的CpG1在普通牦牛中的DNA甲基化状态显著低于多肋牦牛(P<0.05)，普通牦牛的第9和12位CpG位点几乎未甲基化。多肋牦牛中Hoxa10高甲基化在一定程度上抑制了Hoxa10基因的表达，Hoxa6低甲基化水平促进了Hoxa6基因的表达。启动子区域的甲基化在一定程度上影响Hox基因的转录调控，且与多肋性状的形成可能存在一定的联系。

Abstract:

The transcription level and methylation in promoter regions of Hoxa6 and Hoxa10 genes in multi-costa properties and common yak were investigated to explore the mechanism of transcriptional regulation of Hox gene in the process of multi-costa properties.The mRNA expression leves of Hoxa6 and Hoxa10 genes in multi-costa properties and common yak were detected by real-time fluorescence quantitative PCR(qRT-PCR)，and the methylation of Hoxa6 and Hoxa10 genes promoter regions were analyzed by bisulfite-sequencing PCR(BSP)，which included modifying，cloning and sequencing the promoter regions.The results showed that the expression level of Hoxa6 gene was significantly higher in multi-costa properties yak than that in common yak(P<0.05)，while the expression level of Hoxa10 gene was significantly lower in multi-costa properties yak(P<0.05).The methylation status of CpG2 island in the Hoxa6 promoter region was significantly different between multi-costa properties and common yak，especially on No.3，4，8，20 and 21 CpG sites.However，the methylation status of CpG1 island in the Hoxa10 promoter region was significantly higher in multi-costa properties yak than that in common yak(P<0.05)，and there was almost no methylation on No.9 and 12 CpG sites in common yak.All above results suggested that the high DNA hypermethylation of Hoxa10 gene in multi-costa properties yak inhibited the expression of Hoxa10 gene in a certain degree，and the low DNA hypomethylation of Hoxa6 gene enhanced the expression of Hoxa6 gene.It was concluded that methylation in Hox promoter regions may have some influence on transcription regulation of Hox genes，and there may be some correlation with the formation of multi-costa properties.

中图分类号:

熊显荣，张雁，兰道亮，李键，字向东，李善蓉，杨建梅. 金川多肋牦牛Hoxa6和Hoxa10基因甲基化与mRNA差异表达研究[J]. 畜牧兽医学报, 2015, 46(9): 1532-1539.

XIONG Xian-rong，ZHANG Yan，LAN Dao-liang，LI Jian,ZI Xiang-dong，LI Shan-rong，YANG Jian-mei. Differences of Hoxa6 and Hoxa10 Methylation Status and mRNA Expression in Multi-costa Properties of Jinchuan Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(9): 1532-1539.

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金川多肋牦牛Hoxa6和Hoxa10基因甲基化与mRNA差异表达研究

Differences of Hoxa6 and Hoxa10 Methylation Status and mRNA Expression in Multi-costa Properties of Jinchuan Yak

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