摘要： 为鉴定牦牛水通道蛋白7（AQP7）基因，分析其在不同组织中的表达水平及蛋白定位，采用RT-PCR方法克隆九龙牦牛AQP7基因，并进行生物信息学分析，采用RT-qPCR方法检测AQP7基因在九龙牦牛8种不同组织中的表达量，并用免疫组织化学染色方法对AQP7蛋白进行组织表达及定位分析。结果表明，九龙牦牛AQP7基因CDS区序列长度为993 bp，编码330个氨基酸，生物信息学分析发现，AQP7为稳定的疏水性蛋白。进化树结果显示，九龙牦牛AQP7基因与黄牛的亲缘关系最近，同源性为99.7%。AQP7基因在九龙牦牛心脏和肌肉表达量较高，极显著高于肾脏、肝脏、脾脏、肺脏、小肠和瘤胃（P <0.01）。免疫组化结果显示，AQP7蛋白主要分布在九龙牦牛心脏和肌肉的肌细胞以及肾脏近曲小管中，且在心脏、肌肉和肾脏中的表达显著高于肝脏、脾脏、肺脏、小肠和瘤胃（P <0.05）。结果为深入研究AQP7基因在牦牛低氧适应性中的生理功能和能量代谢机制提供了基础数据。

关键词: 九龙牦牛, 水通道蛋白7, 组织表达, 免疫组织化学染色

Abstract: The aim of this study was to identify the Aquaporin 7 gene and analyze the expression level and protein localization of AQP7 gene in different tissues of yak.The AQP7 gene of Jiulong yak was cloned by RT-PCR and analyzed by bioinformatics.RT-qPCR was used to detect the expression levels in 8 different tissues of yaks.Immunohistochemical staining method was used to analyze the expression and location of AQP7 protein in tissues.The results showed that the CDS sequence of AQP7 gene was 993 bp, which encoded 330 amino acids. Bioinformatics analysis found that AQP7 was a stable hydrophobic protein.The phylogenetic tree showed that AQP7 gene of Jiulong yak was closely related to cattle, and its homology was 99.7%. The expression of AQP7 gene was higher in Jiulong yak heart and muscle, which were significantly higher than that in kidney, liver, spleen, lung, intestine and rumen(P <0.01). Immunohistochemical results showed that AQP7 protein was mainly distributed in the muscle cells of the heart and muscles and the proximal convoluted tubules of the kidney, and its expression in the heart, muscles and kidney were higher than that in the liver, spleen, lung, intestine and rumen of Jiulong yak(P <0.05).This study may provide basic data for the physiological function and energy metabolism mechanism in yak high altitude adaptation.

Key words: Jiulong yak, Aquaporin 7, Tissue expression, Immunohistochemistry

中图分类号: 

• Q78

• S823.03