牦牛和犏牛睾丸组织DDX4基因mRNA表达水平与启动子区甲基化

摘要

[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.%[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).