长白猪Plin3、Plin5基因克隆及表达规律研究

脂滴包被蛋白3(Plin3)和脂滴包被蛋白5(Plin5)是细胞内脂滴包被蛋白(PAT)家族的成员,在脂滴合成方面具有重要作用。为探究Plin3和Plin5在长白猪中的序列和表达特征,利用PCR技术扩增该基因,采用生物信息学分析两者的序列特征,并利用实时荧光定量PCR技术检测其在长白猪11个不同组织中的表达特征。结果表明,长白猪Plin3序列全长1 403 bp,Plin5全长1 397 bp,两种蛋白二级结构均以α-螺旋为主,无规则卷曲次之,不存在跨膜结构,无信号肽结构,有多个磷酸化位点。检测两种基因在不同组织中的表达水平发现,Plin3在长白猪的11个组织中均有表达,其中在脾脏中的相对表达量显著高于其他组织(P<0.05),肝脏次之;Plin5在脂肪组织中的相对表达量显著高于其他组织(P<0.05),而在大肠和小肠中不表达。本研究为进一步探究长白猪Plin3和Plin5在脂质代谢中的作用机制提供了理论依据。

Cloning and Expression Pattern of Plin3 and Plin5 Genes in Landraces

Lipid droplet envelope protein 3 (Plin3) and lipid droplet envelope protein 5 (Plin5) are members of the PAT protein family and have important roles in lipid droplet synthesis.To investigate the sequence and expression characteristics of Plin3 and Plin5 in landraces, we used PCR technology to clone and bioinformatics analysis to characterize the sequence characteristics of this two genes. After that, we used real-time fluorescence quantitative PCR to detect their expression characteristics in 11 different tissues of ladraces. The results showed that the full length of Plin3 sequence was 1 403 bp and the full length of Plin5 was 1 397 bp. The secondary structures of both proteins were mainly α-helical, followed by irregularly coiled,without transmembrane structure and signal peptide structure, but multiple phosphorylation sites. The expression levels of both genes in different tissues detected by RT-PCR showed that Plin3 was expressed in 11 tissues of landraces. This paper provides a theoretical basis for further studying the mechanism of Plin3 and Plin5 in fat deposition in landraces.