陆川猪磷酸酪氨酸互作结构域1基因克隆及序列分析

Cloning and Sequence Analysis of PID1 Gene in Luchuan Pig

试验旨在对陆川猪磷酸酪氨酸互作结构域1(phosphotyrosine interaction domain containing 1,PID1)基因进行克隆和生物信息学分析。利用GenBank公布的猪序列设计引物,用RT-PCR扩增得到目的基因片段,并用生物信息学方法分析和预测了陆川猪PID1基因的理化性质与二级结构。结果显示,陆川猪PID1基因编码区全长654bp,编码217个氨基酸;陆川猪PID1基因与莱芜猪、牛、猕猴、人、小鼠、原鸡、大鼠、斑马鱼和非洲爪蟾相对应序列相似性分别为99.08%、87.61%、93.88%、93.58%、90.06%、83.79%、66.94%、66.52%和60.09%。系统进化树分析结果表明,PID1基因在不同物种及进化的过程中具有高度保守性。本研究成功克隆陆川猪PID1基因,为阐明其在陆川猪生长发育及脂肪沉积方面的调控研究奠定了理论基础。

In order to clone the phosphotyrosine interaction domain containing 1 （PID1） gene of Luchuan pig,a pair of special primers was designed according to released sequence of Laiwu pig PID1 gene in GenBank. The PID1 gene was amplified by RT-PCR,its gene sequence characteris- tics and protein structure was systemically analyzed by bioinformatics techniques. The results showed that the cloned PID1 gene fragment included a 654 bp whole length CDS （coding 217 amino acids）. The sequence multi-aligned results showed that Luchuan pig shared 99. 08%, 87.61% ,93.88% ,93.58% ,90.06%,83. 79% ,66.94%,66. 52% and 60.09% of similar nucleotide sequence with that of Laiwu pig, Bos, Macaca, Homos,Mus, Gallus,Rattus, Danio retio and Xenopus, respectively. The phylogenetic tree indicated that PID1 gene was highly conserved in the process of evolution of different species. The cloning and analysis of PID1 gene provided an important foundation for further study regulation mechanism of PID1 gene in Luchuan pig.