静宁鸡PPARα基因克隆与生物信息学分析

Cloning and bioinformatic analysis of PPARα gene in Jingning chicken

为阐明静宁鸡PPARα基因的结构及功能,根据NCBI上登录的原鸡PPARα基因的CDS序列设计引物,以静宁鸡肾脏为材料采用PCR的方法,对目的基因进行克隆,并测序。根据测序的结果,采用各类分析软件,对所获得的DNA片段进行生物信息学分析。结果成功克隆了静宁鸡PPARα基因的全长CDS序列。生物信息学分析结果表明,该基因CDS全长1 407 bp,所编码的蛋白质包含468个氨基酸。其分子量为52 300,等电点为5.85。在二级和三级结构上,α-螺旋和无规则卷曲为蛋白的主要结构形式。PPARα蛋白是一个亲水性蛋白质,不存在跨膜区域,不含信号肽,没有N-糖基化位点,但存在16个O-糖基化位点,存在25个丝氨酸(Ser)磷酸化位点、12个苏氨酸(Thr)磷酸化位点和6个酪氨酸(Tyr)磷酸化位点。静宁鸡PPARα蛋白在第100～168个氨基酸之间有1个ZnF_C4结构域,在第264～449个氨基酸之间有1个HOLI结构域。同源性分析结果表明,静宁鸡PPARα基因与原鸡的亲缘关系最近。静宁鸡PPARα基因的成功克隆和功能预测为进一步研究PPARα基因在静宁鸡脂肪代谢中的作用提供了基础。

In order to clarify the structure and reveal the role of PPARα in Jingning chicken, the CDS sequence of Gallus gallus PPARα was applied, and specific primers were designed accordingly. For the cloning of PPARα, total RNA was extracted from the kidney of Jingning chicken, and PCR was performed. According to the results of sequencing, the obtained DNA fragments were analyzed by various analysis software. Finally, full length CDS sequence of PPARα was successfully obtained. Bioinformatics analysis showed that the CDS of PPARα was 1 407 bp in length, and the encoded protein contained 468 amino acids. Its molecular weight was 52 300 , and the isoelectric point was 5.85. In the secondary structure and tertiary structure, the alpha helix and random coil were the main structural forms of the protein. PPARα was a hydrophilic protein, containing sixteen O -glycosylation sites, twenty-five Ser phosphorylation sites, twelve Thr phosphorylation sites and six Tyr phosphorylation sites. PPARα did not contain transmembrane structure, signal peptides and N -glycosylation sites. The PPARa protein had a ZnF_C4 domain between the 100th and the 168th amino acid, and a HOLI domain between the 264th and the 449th amino acid. The results of homology analysis showed that PPARα gene of Jingning chicken was closely related to Gallus gallus. The successful cloning and functional prediction of PPARα gene in Jingning chicken provide a basis for further study on the role of this gene in the lipid metabolism of Jingning chicken.