摘要： 旨在探究DNA损伤诱导转录本3（DDIT3）基因序列特征及其在母牦牛组织表达特性。以母牦牛为研究对象，利用RT-PCR技术克隆牦牛DDIT3基因，利用生物信息学软件分析DDIT3蛋白的结构和功能，并利用实时荧光定量PCR （RT-qPCR）检测DDIT3基因在牦牛不同组织及不同生理阶段的生殖器官和卵母细胞中的表达特征。结果显示：牦牛DDIT3基因CDS区全长507 bp，共编码168个氨基酸；核苷酸序列比对发现，牦牛与野牛同源性最高，为99.71%，与其他物种的同源性均在88%以上，说明DDIT3基因在进化过程中表现出高度保守性；牦牛DDIT3基因在卵巢中的表达量极显著高于心脏、肝脏、肾脏、脾脏、肺、子宫和输卵管（P <0.01）；在卵巢中，妊娠期DDIT3基因的表达水平显著高于卵泡期、黄体期和胎儿期（P <0.05），黄体期的表达量显著高于胎牛期（P <0.05）；在子宫中，妊娠期DDIT3基因的表达水平显著高于卵泡期和胎牛期（P <0.05）； DDIT3在各时期输卵管中表达水平差异不显著；M Ⅱ期卵母细胞中DDIT3基因的表达水平显著高于GV期和M Ⅰ期（P <0.05），M Ⅱ颗粒细胞DDIT3基因的表达量也显著高于GV期和M Ⅰ期（P <0.05），GV期、M Ⅰ期的卵母细胞和颗粒细胞DDIT3表达量差异不显著。综上，牦牛DDIT3基因可能在维持母牦牛卵巢机能与妊娠以及卵泡发育与成熟过程中发挥重要的调控作用。

关键词: 牦牛, DDIT3基因, RT-qPCR, 组织表达, 卵巢, 卵母细胞

Abstract: The objective of this study was to investigate the sequence and tissue expression characteristics of DNA damage inducible transcript 3(DDIT3) in female yaks. The yak DDIT3 gene was cloned by RT-PCR. The structure and function of DDIT3 protein were analyzed by bioinformatics software. The expression levels of DDIT3 gene in different tissues, and different reproductive organs and oocytes at different physiological stage of female yaks were detected by Real-time quantitative PCR(RT-qPCR). The results showed that the full-length CDS region of yak DDIT3 gene was 507 bp, which encoded 168 amino acids. Nucleotide sequence alignment showed that yak had the highest homology with bison(99.71%) and more than 88% homology with other species, indicating that DDIT3 gene was highly conservative in the process of evolution. The expression of DDIT3 gene in ovary was extremely significantly higher than that in heart, liver, kidney, spleen, lung, uterus and fallopian tube(P <0.01). In the ovary, the expression of DDIT3 gene was significantly higher during pregnancy than that during follicular phase, luteal phase and fetus stage(P <0.05), and it was significantly higher during luteal phase than that in fetus stage(P <0.05). In the uterus, the expression of DDIT3 was significantly higher than that in follicular stage and fetus stage(P <0.05). In oviduct, its expression was not significantly different at different physiological stages. The expression of DDIT3 gene in M Ⅱ stage oocytes was significantly higher than that in GV stage and M Ⅰ stage(P <0.05). Its expression in M Ⅱ granulosa cells was also significantly higher than that in GV stage and M Ⅰ stage(P <0.05). There was no significant difference in DDIT3 expression between oocytes and granulosa cells in GV stage and M Ⅰ stage. In conclusion, yak DDIT3 gene possible plays an important role in regulating ovary function and pregnancy maintenance, follicle development and maturation.

Key words: Bos grunniens, DDIT3 gene, RT-qPCR, Tissue expression, Ovary, Oocyte

中图分类号: 

• S813.1

• Q78