牦牛RETN基因克隆及组织表达分析

Cloning and tissue expression of yak RETN gene

为了克隆RETN基因,并对RETN基因在4.5岁牦牛不同组织器官中的相对表达差异进行分析,提取类乌齐牦牛心脏、肝脏、肺脏、脾脏、臀肌、臀脂、乳腺、大脑组织总RNA,克隆得到RETN基因cDNA片段428 bp,利用RT-qPCR技术检测RETN基因在不同组织器官中的相对表达情况。结果表明:RETN基因开放阅读框长度为330 bp,编码109个氨基酸,与黄牛亲缘关系最近(相似性为99.7%),与小鼠亲缘关系最远(相似性为66.1%);RETN蛋白的理化性质稳定,整条肽链呈亲水性,无信号肽,无跨膜区,主要在线粒体中发挥作用;该蛋白质主要由α-螺旋和无规则卷曲组成;RETN基因在牦牛心脏、肝脏、肺脏、脾脏、臀肌、臀脂、乳腺、大脑组织器官中均有不同程度的表达,在肺脏、脾脏、乳腺组织器官中的相对表达量极显著高于其他组织器官(P<0.01),在臀肌和大脑中的相对表达量较低。说明RETN基因可能在脾脏、肺脏和乳腺中具有调控作用。

This study aimed to clone the RETN gene,and to determine its expression in various tissues,which might lay the foundation for explo-ring the function of RETN gene in 4. 5-year-old yak. The total RNA were extracted from the heart,liver,lung,spleen,gluteal muscle,gluteal fat,mammary gland and brain tissue of the Leiwuqi yak. The 428 bp RETN gene fragment was cloned by RT -qPCR mRNA expressions of RETN among different organs were determined. The results show that the RETN was cloned,its ORF open reading frame is 330 bp in length and encodes 109 amino acids. It has the closest relationship with the cattle and the similarity is 99. 7%. While,the homology with the mouse is the farthest,the similarity is 66. 1%. The physicochemical properties of RETN protein is stable,the whole peptide chain is hydrophilic,no signal peptide,no transmembrane region,mainly plays a role in mitochondria. The secondary structure and tertiary structure analysis results show that the protein is mainly composed of α- Spiral and irregular curl. The RETN gene is widely expressed in the heart,liver,lungs,spleen,gluteus muscle,gluteal fat,mammary gland and brain tissue of the yak. Furthermore,the expression of the gene in the lungs,spleen and mammary gland tissue is extremly higher than that in other tissues ( P<0. 01),and its expression in the gluteal muscles and the brain is lower. It indicates that the RETN gene may have a regulatory role in the spleen,lung and mammary gland in yak.