【摘要】 诱导细胞凋亡的DFF45样效应因子c(cell death-inducing DFFA-like effector c,CIDEC)与脂肪分化密切相关,具有促进脂肪沉积的作用。本研究利用PCR技术克隆了绵羊(Ovis aries)CIDEC基因,并对其序列进行生物信息学分析;采用半定量RT-PCR方法检测了该基因在绵羊主要组织中的表达;同时利用实时荧光定量PCR技术检测了持续饥饿与充足采食状态下阿勒泰羊尾脂CIDEC的差异表达情况。研究结果表明,获得了绵羊CIDEC基因的c DNA序列(Gen Bank登录号:KM199684),其中编码区(coding sequences,CDS)全长714 bp,编码237个氨基酸。经预测CIDEC蛋白存在多个磷酸化位点和蛋白激酶C磷酸化位点。半定量RT-PCR结果显示,CIDEC仅在阿勒泰羊脂肪组织中表达,其中在尾脂中高丰度表达,而在肠系膜脂肪中表达丰度较低。经过持续4周的完全禁食后,CIDEC在阿勒泰羊尾脂组织中表达急剧下降,表达量极显著低于正常充足采食状态(P<0.01)。表明该基因在阿勒泰羊尾脂沉积过程中具有一定的调控作用。研究结果为进一步揭示CIDEC基因在绵羊尾脂组织中的功能提供了参考资料。更多还原

【Abstract】 Cell death-inducing DFFA-like effector c(CIDEC) is related closly to adipose differentiation and promoting fat deposition. In this study, sheep(Ovis aires) CIDEC c DNA was cloned by PCR and analyzed by bioinformatics. Expression profiles of CIDEC in different tissues of sheep were detected by semi-quantitative RT- PCR. The expression levels of CIDEC in tail fat tissue after persistent starvation in Altay sheep was determined by Real-time quantitative PCR(q RT-PCR). The result showed that the coding sequences(CDS) of sheep CIDEC gene was 714 bp and encoded 237 amino acids(Gen Bank accession No.KM199684).Bioinformatic analysis indicated that multiple phosphorylation and protein kinase C sites existed in thesecondary structure of sheep CIDEC protein. The RT-PCR results confirmed that CIDEC m RNA expressed in adipose tissue only, and the expression level was much higher in tail fat than that in esenteric fat. After persistent starvation and fasting for 4 weeks, the expression of CIDEC in tail fat of Altay sheep decreased sharply, its level was very significantly lower than that in normal food-intake sheep(P<0.01). Taken together,these results demonstrated that CIDEC might play an important role in the process of tail fat deposition. In conclusion, this work serves as the basis for further study on CIDEC in the tail fat of sheep.更多还原