摘要：

为明确秦川牛CDS1基因的编码序列、分子特征及其在不同组织的表达特点,以秦川牛为研究对象,利用PCR扩增技术克隆CDS1基因的全编码区序列,利用生物信息学方法分析其全编码区的序列特征,以GAPDH为内参基因,采用qRT-PCR技术检测CDS1基因在脑、睾丸、胰、肺、脾、肾、瘤胃、背最长肌、肝、肌内脂肪和心组织中的表达水平。结果显示,秦川牛CDS1基因编码区为1 392 bp,编码464个氨基酸,CDS1基因在不同物种间具有较高的保守性,且与瘤牛的同源性最高;CDS1基因编码蛋白为疏水性不稳定跨膜蛋白,主要由α-螺旋及不规则卷曲构成;亚细胞定位分析表明,CDS1蛋白主要分布于内质网和线粒体;蛋白互作分析表明,其与PGS1、PPAPDC1系列、CDIPT、PLD系列、CRLS1等与磷脂合成相关的核糖蛋白存在相互作用,提示CDS1基因参与调节磷脂的合成代谢过程;组织表达分析表明,CDS1基因在各个组织均有广泛表达,且在睾丸中表达量最高,在脑的表达水平也较高,二者均显著高于其他组织的表达水平,在心的表达量最低。

关键词: 秦川牛, CDS1基因, 克隆, 生物信息学分析, 组织表达

Abstract:

To clarify the coding sequence and molecular characteristics of the CDS1 gene in Qinchuan cattle and its expression characteristics in different tissues.The full coding region sequence of CDS1 gene was cloned by PCR amplification technique in Qinchuan cattle,and the sequence characteristics of its full coding region were analyzed by bioinformatics.qRT-PCR was used to detect the expression levels of CDS1 gene in brain,testis,pancreas,lung,spleen,kidney,rumen,longest dorsal muscle,liver,intramuscular fat and heart tissues using GAPDH as an internal reference gene.The results showed that the coding region of CDS1 gene in Qinchuan cattle was 1 392 bp,encoding 464 amino acids,and the CDS1 gene was highly conserved among different species and had the highest homology with rumen cattle;the protein encoded by CDS1 gene was a hydrophobic unstable transmembrane protein,mainly composed of α-helix and irregular convolutions,and the subcellular localization analysis showed that CDS1 protein was mainly distributed with endoplasmic reticulum and mitochondria.Protein interaction analysis showed that it interacted with PGS1,PPAPDC1 series,CDIPT,PLD series,CRLS1 and other riboproteins related to phospholipid synthesis,suggesting that CDS1 gene was involved in regulating phospholipid anabolic process;tissue expression analysis showed that CDS1 gene was widely expressed in various tissues,and the highest expression was in testis,and the expression level in brain was also higher.The tissue expression analysis showed that CDS1 gene was widely expressed in all tissues,and its expression level was highest in testis and higher in brain,both of which were significantly higher than that of other tissues,and the expression level was lowest in heart.

Key words: Qinchuan cattle, CDS1 genes, Cloning, Bioinformatics analysis, Tissue expression