猪甲状腺激素应答spot14基因的克隆及组织表达研究

基于电子延伸序列,设计1对克隆引物,成功克隆了猪甲状腺激素应答spot14(thyroid hormone responsive spot14, THRSP)基因(GenBank登录号:JF_951726),并进行了序列分析,同时利用RT-PCR方法分析了THRSP基因的组织分布规律。结果发现,克隆的猪THRSP基因长为621 bp,包括一个完整的开放阅读框架453 bp,编码150个氨基酸。克隆的猪THRSP基因与人、牛、小鼠、褐鼠、欧洲兔、鸡的核苷酸序列同源性分别为83.8%、88.5%、80.9%、80.7%、85.1%、47.1%;推导的cDNA编码区(CDs)的氨基酸序列与人、牛、小鼠、褐鼠、欧洲兔、鸡的同源性分别为76.9%、80.8%、76.2%、76.2%、82.1%、32.0%,构建的系统发育树显示猪与牛的亲缘关系最近。组织表达分析结果表明,THRSP基因在肝脏和脂肪组织中高表达,在脑、脾脏、胃、皮肤、盲肠、直肠中表达量相对较低,在肺脏、十二指肠、空肠、回肠中微量表达,在肾脏、肌肉中无表达。THRSP基因在肝脏、脂肪等脂肪生成组织的高表达暗示了其可能作为调节脂肪合成代谢的调控因子参与脂肪合成,但其调控机理有待进一步研究。

Cloning and Tissue Expression of Sus Scrofa Thyroid Hormone Responsive spot14 Gene

A pair of cloning primers were designed based on the sequence information in silico and were used to clone and analysis the Sus scrofa thyroid hormone responsive spot14(THRSP,accession: JF951726).The positive clone was identified and sequenced,at the same time the THRSP gene organization distribution rule was analyzed by RT-PCR.The length of Sus scrofa THRSP gene was 621 bp,including an open reading flame of 450 bp,encoding 150 amino acids.Identity analysis showed that the THRSP nucleotide sequence in Sus scrofa shared 83.8%,88.5%,80.9%,80.7%,85.1%,47.1% homology with that of Homo sapiens,Bos taurus,Rattus norvegicus,Mus musculus,Oryctolagus cuniculus,Gallus gallus respectively,the predicted peptide shared 76.9%,80.8%,76.2%,76.2%,82.1%,32.0% homology with that of Sus scrofa.The phylogenetic tree demonstrates that Sus scrofa was the closest to cattle.Semi-quantitative RT-PCR analysis showed that the expression level of THRSP was the highest in liver and adipose tissue,the lower in cerebrum,spleen,stomach,skin,caecum and rectum,the lowest in lung,duodena jejunum and ileum,without in kidney and muscle.The high expression level of THRSP in the liver,fat and other fat generation of the organization suggest that which might adjust as fat synthesis of metabolic control factor,but fat synthesis in the regulation mechanism needs further research.