摘要
【目的】了解牦牛和犏牛睾丸组织中DDX4基因mRNA表达水平和启动子区甲基化状态。【方法】采用real-time PCR技术检测牦牛和犏牛睾丸组织DDX4基因mRNA表达水平,采用克隆测序技术获得牦牛和犏牛DDX4基因启动子区序列,采用亚硫酸氢钠测序法检测牦牛和犏牛睾丸组织中DDX4基因启动子区甲基化状态。【结果】牦牛睾丸组织中DDX4基因mRNA表达水平极显著高于犏牛(P<0.01);牦牛和犏牛DDX4基因启动子区1 370 bp,含有核心启动子区(251 bp)和CpG岛(918 bp)。犏牛睾丸组织中DDX4基因启动子区甲基化水平(86.5%)极显著高于牦牛(67.0%)(P<0.01)。【结论】牦牛睾丸组织DDX4基因表达水平极显著高于犏牛,获得了牦牛和犏牛DDX4基因启动子区序列,且犏牛睾丸组织中DDX4基因启动子区甲基化水平极显著高于牦牛(P<0.01)。
[Objective] The study was aimed to investigate the expression levels and promoter methylation status of DDX4 gene in the testes of yak and cattle-yak. [ Method ] Real-time PCR was employed to examine the expression levels in the testes of yak and cattle-yak. Clone sequencing was applied to acquire the promoter sequences of DDX4 gene of yak. The promoter methylation status of DDX4 gene in the testes of yak and cattle-yak was analyzed by using sodium bisulfite sequencing. [Result] The mRNA expression level of DDX4 gene in the testis of yak was significantly higher than that of cattle-yak （P^0.01）. The length of the promoter sequences of the DDX4 gene was 1 370 bp, which included a core promoter （251 bp） and a CpG island （918 bp）. The promoter methylation level of DDX4 gene in the testes of cattle-yaks （86.5%） was significantly higher than that in yak （67.0%） （P〈0.01）. [Conclusion] Results of the study showed that the testicular expression level of DDX4 might be involved in cattle-yak male sterility, and the promoter hypermethylation was correlated to the lower expression of DDX4 in the testes of cattle-yak.
出处
《中国农业科学》
CAS
CSCD
北大核心
2013年第3期630-638,共9页
Scientia Agricultura Sinica
基金
国家自然科学基金(30500360)
农业生物技术国家重点实验室开放课题(2009SKLAB07-2)
