中华鳖gp130基因全长cDNA克隆及生物信息学分析

Molecular Cloning and Bioinformatics Analysis of gp130 in Chinese Soft-Shelled Turtle(Pelodiscus sinensis)

目的:从中华鳖脾脏、肝脏、肠道构建的SMART cDNA文库中克隆中华鳖gp130基因cDNA全长。方法:采用RACE法克隆中华鳖gp130基因全长cDNA,并对其进行生物信息学分析。结果:中华鳖gp130基因cDNA全长3806 bp,对应基因组全长73 252 bp,包含16个内含子及17个外显子。中华鳖gp130与鸟类、哺乳类相比均有保守的共线性关系。该基因编码由927个氨基酸残基构成的序列,二级结构主要包括α螺旋、延伸链、无规则卷曲、β转角,三级结构包含配体结合、跨膜结构域、纤维链接蛋白结构域等。系统进化分析显示与鸟类首先聚类,其次是哺乳类,最后是两栖类与鱼类。同时以人GP130蛋白为模型构建了中华鳖GP130蛋白的3D结构模型,一致性为58.32%。结论:获得了中华鳖gp130基因全长并做了生物信息学分析,为深入研究GP130及相关信号通路提供了基础。

Objective： To clone the gp130 full-length cDNA from Chinese soft-shelled turtle.Methods： RACE PCR and bioinformation were used to clone the full-length sequence and predict the gp130 gene and protein. Result： The length of gp130 cDNA of Chinese soft-shelled turtle was 3806 bp, encoding a 927 amino acid sequence and its genomic DNA length was 73 252 bp, which spans 16 introns and 17 exons. The BLAST results suggested that the gp130 is a homolog of those of birds and mammalians. Bioinformatics analysis showed that the secondary structure consists of α-helix, extension strand, random coil and β-turn. The result of tertiary structure analysis showed that the protein contains ligand binding domain, transmembrane domains and FN3 domains. Phylogenetic analysis demonstrated that the reptile gp130 clustered firstly with birds, and secondly with mammals, and finally with amphibians and fish. Meantime, we use the human GP130 protein as a model to construct the turtle GP130, the identity was 58.32%.Conclusion： We cloned the gp130 full-length cDNA from Chinese soft-shelled turtle. This study laid a basement for further study of gp130 and the related signal pathways.