山羊CIDE家族基因克隆分析及互作蛋白预测鉴定

Cloning and Analysis of Goat CIDEs and Identification of Predictive Interaction Proteins

旨在获得CIDEB、CIDEC基因的CDS,检测该家族基因在山羊不同组织中的表达量,预测其互作蛋白,为进一步揭示CIDE家族基因在脂代谢中的调控网络提供参考。主要利用RT-PCR克隆获得山羊CIDEB、CIDEC基因序列并利用在线工具分析CIDE家族蛋白的生物学特性,并预测其互作蛋白。利用实时荧光定量PCR(RT-qPCR)检测了CIDE家族基因在山羊不同组织中的表达水平,并检测各互作蛋白在其表达水平最高的组织中的表达量,利用SPSS分析其表达量之间的相关性。克隆获得了CIDEB基因CDS区660 bp,共编码219个氨基酸,CIDEC基因CDS区714 bp,编码237个氨基酸。进化树分析显示,山羊CIDEA、CIDEB、CIDEC均与绵羊亲缘关系最近。RT-qPCR检测发现CIDEA、CIDEB、CIDEC基因分别在山羊瘤胃、肝脏、小肠中表达量最高。CIDEA与DFFB和ELOVL3在瘤胃中的表达量具有极显著相关性(P<0.01),与DIO2、TMEM26、PRDM16、PLIN1具有显著相关性(P<0.05)。CIDEB与DFFA和SDR39U1在肝脏中的表达量具有显著相关性(P<0.05)。CIDEC与PLIN2、GPLD1、ADIPOQ在小肠中的表达量具有显著相关性(P<0.05)。通过分析发现了与CIDE家族表达量具有相关性的基因,可为进一步揭示CIDE基因在脂质代谢中的作用及其调控网络提供理论基础,且CIDE家族蛋白均为脂滴相关蛋白,也可为进一步研究精确的脂滴形成机制提供参考。

The aim of this study was to obtain the CDS of CIDEB and CIDEC gene,detect the expression of CIDEB and CIDEC in different tissues of goats,predict their interaction proteins.It would provide basial data for revealing the regulatory role of CIDEs in lipid metabolism.The CDS of CIDEB and CIDEC gene were cloned by RT-PCR,and the biological characteristics and interaction proteins of CIDE proteins were analyzed by online tools.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of CIDEs and the genes,encoding the interaction proteins,in different tissues of goats,following the correlation analyses by a SPSS software.The CDS of CIDEB was 660 bp,encoding 219 amino acids,while the CDS of CIDEC was 714 bp,encoding 237 amino acids.Phylogenetic tree analysis showed that goat CIDEA,CIDEB and CIDEC were closely related to corresponding genes from sheep.RT-qPCR detection showed that CIDEA,CIDEB and CIDEC were highly expressed in the rumen,liver and small intestine of goat,respectively.The expression of CIDEA was significantly correlated with the expression of DFFB(P<0.01),ELOVL3(P<0.01),DIO2(P<0.05),TMEM26(P<0.05),PRDM16(P<0.05)and PLIN1(P<0.05)in rumen.The expression of CIDEB was significantly correlated with that of DFFA and SDR39U1 in liver(P<0.05).The expression of CIDEC was obviously correlated with that of PLIN2,GPLD1 and ADIPOQ in small intestine(P<0.05).Through the analysis,we found genes related to the expression level of CIDEs,which could provide a theoretical basis for further revealing the role of CIDE gene in lipid metabolism and its regulatory network.Moreover,the proteins of CIDEs were all lipid-droplet related proteins,and could also provide a reference for further studying the precise mechanism of lipid droplet formation.