梅花鹿CAV1基因cDNA克隆及序列分析

cDNA cloning and sequence analysis of CAV1 gene in Cervus nippon

为了为深入研究梅花鹿窖蛋白-1(Caveolin-1,CAV1)基因的生物学功能提供数据支持,试验利用RT-PCR和分子克隆技术获得梅花鹿CAV1基因CDS序列并对其进行生物信息学分析。结果表明:梅花鹿CAV1基因序列与马鹿、山羊相似度较高,与加拿大猞猁、智人等相似度较低;CDS区长537 bp,编码178个氨基酸;CAV1蛋白是Caveolin家族的一员,在第97~119位存在1个跨膜螺旋区,为疏水性稳定酸性蛋白,无信号肽;该蛋白中α-螺旋和无规则卷曲占比较高,亚细胞定位主要在细胞膜;CAV1蛋白有13个磷酸化位点,1个乙酰化位点;CAV1蛋白参与细胞内吞作用、黏着斑、细菌侵袭上皮细胞等通路;该蛋白与CAV2、NOS3、EGFR、FYN等蛋白存在相互作用。

In order to provide data support for further study on the biological function of Caveolin-1(CAV1)gene in sika deer,the CDS sequence of CAV1 gene was obtained by RT-PCR and molecular cloning technology,and bioinformatics analysis was performed.The results showed that the CAV1 gene sequence of Sika deer had a high similarity with that of red dear(Cervus Canadensis)and goat(Capra hircus),however,low with Canadian lynx(Lynx Canadensis)and man(Homo sapiens).The CDS sequence of CAV1 gene is 537 bp long,encoding 178 amino acids.The CAV1 protein,a member of Caveolin family,is known as hydrophobic and acid-stable protein with a transmembrane helical region in the 97-119 position,without signal peptide.Two types of secondary structure ofα-helix and random coil were highly regular in the CAV1 protein which can be found the most in cell membrane.Thirteen phosphorylation sites and one acetylation sites were found in the primary structure of CAV1 protein.CAV1 protein is involved in endocytosis,focal adhesion,bacterial invasion of epithelial cells and other pathways.Moreover,the Cav1 proteins were found to be able to interact with CAV2,NOS3,EGFR,FYN and other proteins.