绵羊RBP4基因克隆及其在不同月龄睾丸中的表达

发布时间：2018-07-10 03:49

本文选题：睾丸 + 绵羊　；参考：《农业生物技术学报》2017年05期

【摘要】：本研究旨在克隆绵羊(Ovis aires)视黄醇结合蛋白4(retinol binding protein 4,RBP4)基因的CDS区序列,检测其在绵羊不同月龄睾丸组织中的表达水平,从而为研究RBP4基因在绵羊睾丸发育中的作用奠定基础。以绵羊睾丸为材料,采用RT-PCR技术克隆RBP4基因并用相关生物信息学软件预测RBP4蛋白的结构,采用qRT-PCR和免疫组织化学技术分别检测RBP4 m RNA和蛋白质的表达水平。结果显示,克隆得到绵羊RBP4基因CDS区长度为606 bp,编码201个氨基酸。绵羊RBP4核苷酸序列与山羊(Capra hircus)、牛(Bos taurus)、猪(Sus scrofa)、人(Homo sapiens)和小鼠(Mus musculus)的相似性分别为97%、97%、95%、91%和85%。绵羊RBP4基因与山羊具有最近的亲缘关系,其次为牛、猪、人和小鼠。RBP4 m RNA在不同月龄绵羊睾丸中均有表达,表达量随月龄增加呈现出先降低后上升的趋势,相邻两组之间差异极显著(P0.01)。RBP4蛋白主要表达于初级精母细胞、精子细胞、精子和生精小管,阳性反应积分光密度值随月龄增加呈现出先升高后降低的趋势,与绵羊睾丸RBP4 m RNA表达量相反。本研究结果表明,RBP4基因可能参与绵羊睾丸发育的调控,为进一步探讨RBP4在绵羊生殖过程中发挥的作用提供参考资料。
[Abstract]:The aim of this study was to clone the CDS region of the retinol binding protein 4 (RBP4) gene of sheep (vis aires) and detect its expression level in the testis of sheep at different months of age, so as to lay a foundation for the study of the role of RBP4 gene in the development of sheep testis. RBP4 gene was cloned from sheep testis by RT-PCR and the structure of RBP4 protein was predicted by bioinformatics software. The expression levels of RBP4 mRNA and protein were detected by qRT-PCR and immunohistochemistry, respectively. The results showed that the length of CDS region of sheep RBP4 gene was 606 BP, encoding 201 amino acids. The similarity between sheep RBP4 nucleotide sequence and goat (Bos taurus), pig (Sus scrofa), human (Homo sapiens) and mouse (Mus musculus) was 97,9795% and 85.1%, respectively. Sheep RBP4 gene was closely related to goat, followed by bovine, pig, human and mouse. RBP4 mRNA was expressed in testis of sheep at different months of age. The expression of RBP4 mRNA decreased first and then increased with the increase of age. The difference between the two groups was significant (P0.01). RBP4 protein was mainly expressed in primary spermatocytes, spermatozoa and seminiferous tubules. The integral optical density of positive reaction increased first and then decreased with the increase of age. The expression of RBP4 mRNA was opposite to that of sheep testis. The results suggest that RBP4 gene may be involved in the regulation of testis development in sheep, and provide references for further study of the role of RBP4 in sheep reproductive process.
【作者单位】：甘肃农业大学动物科学技术学院;甘肃肉羊繁育生物技术工程实验室;甘肃农业大学动物医学院;
【基金】：甘肃省科技重大专项计划(No.1602NKDH020) 甘肃农业大学校级课题(No.GSAU-ZL-2015-030) 甘肃省农业生物技术研究与应用开发项目(No.GNSW-2012-19) 国家肉羊产业技术体系(No.CARS-39)
【分类号】：Q78;S826

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